The Junk DNA Wars

This month, the New York Times' published a report on “the junk DNA wars” asking “Is Most of Our DNA Garbage”? 1/ Readers of the article (and an anonymous follow-up piece on the reactions appearing in science blogs) 2/ would come away thinking that there is a serious debate in the scientific community over the proposition that “junk DNA” is “mostly functional.”

Without defining terms like “functional” and “junk,” however, it is impossible to know what is in dispute and what is not.The follow-up piece is particularly frustrating. It observes that

Some scientists, like T. Ryan Gregory, a evolutionary biologist ... argue that if DNA is mostly functional, then it’s hard to explain why rather humble species, like the onion, have far more DNA than we do. ...

Those who disputed Gregory’s findings [sic — Gregory did not discover the long-standing C-value paradox 3/ ], including supporters of intelligent design, cited the Encode Project, an N.I.H.-sponsored attempt to catalog the functional elements of the genome. Encode scientists found that 80 percent of the genome had “biochemical functions,” suggesting that there was a lot less junk DNA than scientists had thought. But did “biochemical function” really mean anything?

For many scientists, it didn’t. A University of Toronto biochemist, Larry Moran, wrote that “the general public has been snowed by the Encode publicity campaign and by naïve journalists who have enthusiastically reported that junk DNA is dead.”

But the Times' writers did not explain why “many scientists” are not snowed by the 80% statistic. After reading some of the ENCODE papers and the surrounding (typically hyperbolic) publicity, I concluded that:

The ENCODE papers show that 80% of the genome displays signs of certain types of biochemical activity—even though the activity may be insignificant, pointless, or unnecessary. This 80% includes all of the introns, for they are active in the production of pre-mRNA transcripts. But this hardly means that they are regulatory or otherwise functional. Indeed, if one carries the ENCODE definition to its logical extreme, 100% of the genome is functional—for all of it participates in at least one biochemical process—DNA replication.

That the ENCODE project would not adopt the most extreme biochemical definition is understandable—that definition would be useless. But the ENCODE definition is still grossly overinclusive from the standpoint of evolutionary biology. From that perspective, most estimates of the proportion of “functional” DNA are well under 80%. 4/

In short, evolutionary biologists reject "biochemical function" as a criterion for recognizing "junk" because not every bit of biochemical activity affects the reproductive fitness of organisms. (Neither does chemical activity per se show any influence on phenotypes that are related to the healthy functioning of those organisms.) To the evolutionary biologists, the term “junk DNA” means parts of the genome in which the particular DNA sequences (the order of the base pairs) do not have evolutionary significance. The Times article defines “junk DNA” differently, and vaguely, as “pieces of DNA that do nothing for us.” This is not the scientific definition. In fact, the earliest papers on “junk DNA” proposed that much of it might “do something” for us.

The “junk DNA war” (or rather the confusion about the meaning of the term “junk”) has spilled over into the legal realm. A brief that leading genetics and genomics researchers submitted to the U.S. Supreme Court to clarify the privacy implications of forensic DNA typing tried to address it. 5/ These researchers observed that

• In genetics, “junk DNA” denotes sequences that lie outside of genes and that are not under detectable selective pressure: that such DNA exists is not in doubt.
• “Junk” DNA sequences could be biologically useful or interesting yet not be useful for disease diagnosis or prediction.
• ENCODE data do not reveal that anywhere near 80% of the genome contains medically relevant information.
• The ENCODE findings indicate that the system that regulates gene expression is exquisitely complex, but they do little to change the status of “junk DNA” in general.

As far as I know, these conclusions have not been contradicted by new studies, but I have not conducted a recent literature review and would be grateful to hear of relevant papers that undermine these observations.

Notes

1. Carl Zimmer, Is Most of Our DNA Garbage?, N.Y. Times Mag., Mar. 5, 2015 
2. Re: Is Most of Our DNA Garbage?, N.Y. Times Sunday Mag., Mar. 20, 2015
3. See Sean R. Eddy, The C-value Paradox, Junk DNA and ENCODE, 22 Current Biology R898 (2012)
4. David H. Kaye, ENCODE’S “Functional Elements” and the CODIS Loci (Part II. Alice in Genomeland), Forensic Science, Statistics, and the Law, Sept. 18, 2012 (note omitted)
5. Brief of Genetics, Genomics, and Forensic Science Researchers as Amici Curiae in Support of Neither Party, Maryland v. King, No. 12-204, Dec, 28, 2012, reprinted in part in Henry T. Greely & David H. Kaye, A Brief of Genetics, Genomics and Forensic Science Researchers in Maryland v. King, 53 Jurimetrics J. 43 (2013), available at http://ssrn.com/abstract=2403063http://ssrn.com/abstract=2403063. Disclosure statement: I prepared an initial draft of the brief and coordinated the revisions to it.

"Scientists' Brief" on CODIS Loci: Q & A

On November 9, 2012, the Supreme Court voted to review a case posing the following question: “Does the Fourth Amendment allow the States to collect and analyze DNA from people arrested and charged with serious crimes?” In Maryland v. King, the state’s supreme court concluded that the protection against unreasonable searches and seizures forbids the state from collecting DNA from an individual whose true identity can be established with ordinary fingerprints. On December 28, 2012, the Supreme Court received a Brief of Genetics, Genomics and Forensic Science Researchers as Amici Curiae. Below are several questions and answers about the brief.

Who contributed to the brief?

I did, and Hank Greely was an additional author. The scientists who participated in the writing are all active and distinguished researchers at medical schools (including Harvard, Yale, and Johns Hopkins) or universities (including Duke, Penn State, and Kings College, London). They include a former president of the American Society of Human Genetics, a past president of the American Board of Medical Genetics, Fellows of the American Association for the Advancement of Science, and members of the Institute of Medicine and the American Academy of Arts and Sciences.

Why did these law professors, medical and statistical geneticists, and molecular biologists submit an amicus brief?

The brief is intended “to inform the Court of the possible medical and social significance of the DNA data stored in law enforcement databases.” (P. 1). Advocacy groups, legal scholars, and some judges have asserted that the small number of features used in law enforcement DNA databases are predictive of health status (or soon will be). The brief attempts to clarify this issue.

Which side does the brief support?

The brief was submitted in support of neither side. It describes the nature of genetic information, the features of the genome used in law enforcement DNA databases, how those features are used in medical research, and whether they currently permit police, employers, or insurers to discern significant facts about a person’s present or future health status.

What conclusions does it reach?

Amici conclude that “[u]nlike medical genetic tests, law enforcement identification profiles have no known value for medical diagnosis or prediction of future health.” (P. 2).

That’s today. What about the future?

Amici caution that “no one can say with certainty what the future will bring, and it is possible that specific loci will be found to affect the operation of certain genes or to display correlations to disease states.” (P. 2). Nevertheless, they suggest that “it is unlikely that the identification profiles will turn into powerful medical diagnostic or predictive tools that can be used to infer disease states or predispositions by examining forensic database records.” (P.2).

Does this mean that the “CODIS loci,” as the identifying features are called, have no medical significance?

Absolutely not. The DNA sequences have been used in medical research for some 20 years to hunt for disease-causing gene mutations. They have been studied for associations with diseases and traits such as longevity. The question the brief addresses is what kind of information can be gleaned from inspecting a database record.

Doesn’t the highly publicized ENCODE Project prove that there is no such thing as “junk DNA”?

The brief contends that “debate over the fraction of the genome that is, in an evolutionary sense, 'junk' ... is orthogonal to the matter before the Court.” (P. 26). A section of the brief explains that the data sets and papers recently released from the international Encyclopedia of DNA Elements Project are important to further research into gene regulation and other matters, but they do not indicate that all DNA sequences are critical to health or other important traits. What “[t]he ENCODE papers show [is] that 80% of the genome displays signs of certain types of biochemical activity—even though the activity may be insignificant, pointless, or unnecessary.” (P. 32).

Well, how about other uses? Don’t the CODIS loci tell scientists a lot about a person’s ancestry and race?

Not really. The CODIS loci can reveal something about bio-geographic ancestry, but anthropologists and population geneticists use far more probative ancestry-informative and lineage markers to study genetic histories. That “race” is not a biological category is now well known. As for socially perceived race, “[a] CODIS profile could be used to calculate probabilities that someone would be described as Caucasian, African-American, or Hispanic, but categorical inferences would not be very accurate, and attempts to predict the census-type race of a person from a CODIS profile would seem pointless considering that apparent race already would be known.” (P. 36).

So the brief shows that there is absolutely no important information that can be deduced from a CODIS profile?

No, amici do not say that either. The brief explains that “[b]ecause children inherit all their DNA from their biological parents, the CODIS loci can be powerful tools for determining whether two people could be genetically related as parent and child. ... [T]he most powerful genetic information other than identity that the CODIS profiles contain [would be] that two people are not parent and child” or “that two people were identical twins.” (Pp. 33-34).

Where can I find the brief?

Here is a pdf. It also should appear."soon," along with other briefs, on the American Bar Association's Preview of Supreme Court cases.

Postscript: The brief and an introduction to it is published as Henry T. Greely & David H. Kaye, A Brief of Genetics, Genomics and Forensic Science Researchers in Maryland v. King, 53 Jurimetrics J. 43 (2013). The publication is available at http://ssrn.com/abstract=2403063

Dear Judges: A Letter from the Electronic Frontier Foundation to the Ninth Circuit

On the eve of the en banc oral argument in Haskell v. Harris, The Electronic Frontier Foundation (EFF) filed a letter asking "the Court to consider the ENCODE project findings in determining the outcome of this case." It seems hard to oppose the idea that the court should consider relevant scientific research, but without input from the scientific community, will the judges do better than they have in the past as "amateur scientists" (to use the skeptical phrase of Chief Justice Rehnquist in Daubert v. Merrell Dow Pharmaceuticals, Inc.)?

Deciphering the ENCODE papers' descriptions of the data is no easy task, and EFF's lawyers do not seem to be up to it. Their letter asserts that the project "has determined that more than 80% of DNA once thought to be no more than 'junk' has at least one biochemical function, controlling how our cells, tissue and organs behave." This is not a fair characterization of the findings. Which geneticist ever claimed that all noncoding DNA plays no role in how cells behave? The issue always has been how much junk, how much func -- and what "functions"?

What does EFF mean by "controlling"? Making organs function? Stimulating tissue growth? Turning normal cells into cancerous ones? Making us tall or short, fat or skinny, gay or straight? None of those things are mentioned in the Nature cover story cited in the letter. Instead, the EFF relies on New York Times reporter Gina Kolata's misleading news article for EFF's claim that "The ENCODE project has determined that 'junk' DNA plays a critical role in determining a person’s susceptibility to disease and physical traits like height."

My earlier postings described the limited meaning of the phrase "biochemical function" in the cited paper. I'd love to see a citation to a page of an ENCODE paper that asserts that fully 80% of the noncoding DNA is determining "susceptibility to disease and physical traits like height." And if I were a judge, I would demand an explanation of why "physical traits like height" are, in the words of the EFF letter, "sensitive and private."

After the judges consider the ENCODE papers (by having their law clerks read them?), will they be better informed about the actual privacy implications of the CODIS loci than they were before this excursion into this realm of the bioinformatics? I would not bet on it, but maybe I am growing cynical.

ENCODE’S “Functional Elements” and the CODIS Loci (Part II. Alice in Genomeland)

Yesterday, I introduced the concepts and terms required to ascertain whether the estimated proportion of the genome that encodes the structure of proteins or regulates gene expression has jumped from 5 or 10% to 80%. Today, I shall focus on the possible meanings of "functional" to show that this is not what the ENCODE papers state or imply. “Functional” is an adjective, and Alice learned from Humpty Dumpty that adjectives are malleable:

"When I use a word," Humpty Dumpty said, in rather a scornful tone, "it means just what I choose it to mean—neither more nor less."
"The question is," said Alice, "whether you can make words mean so many different things."
"The question is," said Humpty Dumpty, "which is to be master—that's all."
Alice was too much puzzled to say anything, so after a minute Humpty Dumpty began again. "They've a temper, some of them—particularly verbs, they're the proudest—adjectives you can do anything with, but not verbs—however, I can manage the whole lot! Impenetrability! That's what I say!"

Like Humpty, who was redefining the word “glory,” the ENCODE authors recognized that “functional” can have many meanings. As Ewan Birney later explained:

Like many English language words, “functional” is a very useful but context-dependent word. Does a “functional element” in the genome mean something that changes a biochemical property of the cell (i.e., if the sequence was not here, the biochemistry would be different) or is it something that changes a phenotypically observable trait that affects the whole organism?^1/

Still other possibilities exist. For example, the first paper to use the adjective “junk” for noncoding DNA noted that even debris accumulated in the course of evolution or introduced from viral infections could have a function simply by creating spaces between genes.^2/ The pieces of dead wood that are joined together to form the hull of a row boat have a function—they exclude the water from the vessel to keep it afloat. This does not mean that the detailed structure of the planks—the precise width of each plank or the number of ridges on its surface—affects its functionality. And, just as something can be inactive and functional, so too something can be alive with activity and yet be nonfunctional.

ENCODE uses biochemical activity—the notion that “the biochemistry would be different”—as a synonym for functional. Here is the definition of “functional” in the top-level paper:

Operationally, we define a functional element as a discrete genome segment that encodes a defined product (for example, protein or non-coding RNA) or displays a reproducible biochemical signature (for example, protein binding, or a specific chromatin structure).^3/

This definition may be useful for the purpose of describing the size of ENCODE’s catalog of elements for later study, but it contrasts sharply with the notion of functional as affecting a nontrival phenotype. The ENCODE papers show that 80% of the genome displays signs of certain types of biochemical activity—even though the activity may be insignificant, pointless, or unnecessary. This 80% includes all of the introns, for they are active in the production of pre-mRNA transcripts. But this hardly means that they are regulatory or otherwise functional.^4/ Indeed, if one carries the ENCODE definition to its logical extreme, 100% of the genome is functional—for all of it participates in at least one biochemical process—DNA replication.

That the ENCODE project would not adopt the most extreme biochemical definition is understandable—that definition would be useless. But the ENCODE definition is still grossly overinclusive from the standpoint of evolutionary biology. From that persective, most estimates of the proportion of “functional” DNA are well under 80%. Various biologists or related specialists have provided varying guestimates:

• Under 50%: “About 1% … is coding. Something like 1-4% is currently expected to be regulatory noncoding DNA ... . About 40-50% of it is derived from transposable elements, and thus affirmatively already annotated as “junk” in the colloquial sense that transposons have their own purpose (and their own biochemical functions and replicative mechanisms), like the spam in your email. And there’s some overlap: some mobile-element DNA has been co-opted as coding or regulatory DNA, for example. [¶] … Transposon-derived sequence decays rapidly, by mutation, so it’s certain that there’s some fraction of transposon-derived sequence we just aren’t recognizing with current computational methods, so the 40-50% number must be an underestimate. So most reasonable people (ok, I) would say at this point that the human genome is mostly junk (“mostly” as in, somewhere north of 50%).”^5/


• 40%: “ENCODE biologist John Stamatoyannopoulos … said … that some of the activity measured in their tests does involve human genes and contributes something to our human physiology. He did admit that the press conference mislead people by claiming that 80% of our genome was essential and useful. He puts that number at 40%.”^6/


• 20%: “[U]sing very strict, classical definitions of “functional” [to refer only to] places where we are very confident that there is a specific DNA:protein contact, such as a transcription factor binding site to the actual bases—we see a cumulative occupation of 8% of the genome. With the exons (which most people would always classify as “functional” by intuition) that number goes up to 9%. … [¶] In addition, in this phase of ENCODE we did [not] sample … completely in terms of cell types or transcription factors. [W]e’ve seen [at most] around 50% of the elements. … A conservative estimate of our expected coverage of exons + specific DNA:protein contacts gives us 18%, easily further justified (given our [limited] sampling) to 20%.”^7/

So why did the ENCODErs opt for the broadest arguable definition? Birney’s answer is that it describes a quantity that the project could measure; that the larger number underscores that a lot is happening in the genome; that it would have confused readers to receive a range of numbers; and that the smaller number would not have counted the efforts of all the researchers.

Whether these are very satisfactory reasons for trumpeting a widely misunderstood number is a matter that biologists can debate. All I can say is that (1) I have been unable to extract a clear number—whatever one should make of it—for a percentage of the genome that constitutes the regulatory elements—the promoters, enhancers, silencers, ncRNA “genes,” and so on; (2) this number is almost surely less than the 80% figure that, at first glance, one might have thought ENCODE was reporting; and (3) “functional element” as defined by the ENCODE Project is not a term that has clear or direct implications for claims of the law enforcement community that the loci used in forensic identification are not coding and therefore not informative.

Of course, none of this means that the description from law enforcement is correct. It simply means that even after this phase of ENCODE, there are still a huge number of base pairs that might or might not be regulatory or influence regulation and hence, gene expression. And the CODIS STRs might or might not be among them. Published reports suggest that they are not,^8/ but the logic that just because a DNA sequence is noncoding (and nonregulatory), it conveys zero information about phenotype is flawed. It overlooks the possibility of a correlation between the nonfunctional sequence (because it sits next to an exon or a regulatory sequence).^9/ Again, however, the published literature reviewing the CODIS STRs does not reveal any population-wide correlations that permit valid and strong inferences about disease status or propensity or other socially significant phenotypes.^10/

Will this situation change? A thoughtful answer would take up a lot of space.^11/ For now, I'll just repeat the aphorism attributed to Yogi Berra, Neils Bohr, and Storm P: "It's hard to make predictions, especially about the future."

Notes

1. Ewan Birney, ENCODE: My Own Thoughts, Ewan’s Blog: Bioinformatician at Large, Sept. 5, 2012, http://genomeinformatician.blogspot.co.uk/2012/09/encode-my-own-thoughts.html.

2. David E. Comings, The Structure and Function of Chromatin, in 3 Advances in Human Genetics 237, 316 (H. Harris & K. Hirschhorn eds. 1972) (“Large spaces between genes may be a contributing factor to the observation that most recombination in eukaryotes is inter- rather than intragenic. Furthermore, if recombination tended to be sloppy with most mutational errors occurring in the process, it would an obvious advantage to have it occur in intergenic junk.”). For more discussion of this paper, see T. Ryan Gregory, ENCODE (2012) vs. Comings (1972), Sept. 7, 2012, http://www.genomicron.evolverzone.com/2012/09/encode-2012-vs-comings-1972/.

3. Ian Dunham et al., An Integrated Encyclopedia of DNA Elements in the Human Genome, 489 Nature 57 (2012).

4. These regions do contain some RNA-coding sequences, and those small parts could be doing something interesting (producing RNAs that are regulatory or that defend against infection by viral DNA, for example), but this kind of activity does not exist in the bulk of the introns that are, under the ENCODE definition, 100% functional.

5. Sean Eddy, ENCODE Says What?, Sept. 8, 2012, http://selab.janelia.org/people/eddys/blog/?p=683. He adds that:

[A]s far as questions of “junk DNA” are concerned, ENCODE’s definition isn’t relevant at all. The “junk DNA” question is about how much DNA has essentially no direct impact on the organism’s phenotype—roughly, what DNA could I remove (if I had the technology) and still get the same organism. Are transposable elements transcribed as RNA? Do they bind to DNA-binding proteins? Is their chromatin marked? Yes, yes, and yes, of course they are—because at least at one point in their history, transposons are “alive” for themselves (they have genes, they replicate), and even when they die, they’ve still landed in and around genes that are transcribed and regulated, and the transcription system runs right through them.

6. Faye Flam, Skeptical Takes on Elevation of Junk DNA and Other Claims from ENCODE Project, Sept. 12, 2012, http://ksj.mit.edu/tracker/2012/09/skeptical-takes-elevation-junk-dna-and-o. Stamatoyannopoulos added that:

What the ENCODE papers … have to say about transposons is incredibly interesting. Essentially, large numbers of these elements come alive in an incredibly cell-specific fashion, and this activity is closely synchronized with cohorts of nearby regulatory DNA regions that are not in transposons, and with the activity of the genes that those regulatory elements control. All of which points squarely to the conclusion that such transposons have been co-opted for the regulation of human genes -- that they have become regulatory DNA. This is the rule, not the exception.

7. Ewan Birney, ENCODE: My Own Thoughts, Ewan’s Blog: Bioinformatician at Large, Sept. 5, 2012, http://genomeinformatician.blogspot.co.uk/2012/09/encode-my-own-thoughts.html.

8. E.g., Sara H. Katsanis & Jennifer K. Wagner, Characterization of the Standard and Recommended CODIS Markers, J. Forensic Sci. (2012).

9. E.g., David H. Kaye, Two Fallacies About DNA Databanks for Law Enforcement, 67 Brook. L. Rev. 179 (2001).

10. E.g., Sara H. Katsanis & Jennifer K. Wagner, Characterization of the Standard and Recommended CODIS Markers, J. Forensic Sci. (2012); Jennifer K. Wagner, Reconciling ENCODE and CODIS, Penn Medicine News Blog, Sept. 18, 2012, http://news.pennmedicine.org/blog/2012/09/reconciling-encode-and-codis.html.

11. For my earlier, and possibly dated, effort to evaluate the likelihood that the CODIS loci someday will prove to be powerfully predictive or diagnostic, see David H. Kaye, Please, Let's Bury the Junk: The CODIS Loci and the Revelation of Private Information, 102 Nw. U. L. Rev. Colloquy 70 (2007) and Mopping Up After Coming Clean About "Junk DNA", Nov. 23, 2007.

ENCODE’S “Functional Elements” and the CODIS Loci (Part I)

Last week I noted some of the hyperbolic headlines accompanying the coordinated publication of a large number of datasets from the ENCODE Project . The abstract of the top-level paper begins as follows:

The human genome encodes the blueprint of life, but the function of the vast majority of its nearly three billion bases is unknown. The Encyclopedia of DNA Elements (ENCODE) project has systematically mapped regions of transcription, transcription factor association, chromatin structure and histone modification. These data enabled us to assign biochemical functions for 80% of the genome, in particular outside of the well-studied protein-coding regions.^1/

Hoping to decipher these sentences, I have been reading about gene regulation. This modest effort stems from more than academic curiosity. If the popular and even some of the scientific press is to be believed, ENCODE has exorcized “junk DNA” from the body of scientific knowledge.^2/ The bright light suddenly shining on the “dark matter” of the genome (to introduce another sloppy metaphor)^3/ raises a giant question mark for the criminal justice system. Law enforcement authorities have always insisted that the snippets of DNA used to generate DNA identification profiles are just nonfunctional "junk."^4/ Now, according to New York Times science correspondent Gina Kolata,

As scientists delved into the “junk” — parts of the DNA that are not actual genes containing instructions for proteins — they discovered a complex system that controls genes. At least 80 percent of this DNA is active and needed. … [¶] … The thought before the start of the project, said Thomas Gingeras, an Encode researcher from Cold Spring Harbor Laboratory, was that only 5 to 10 percent of the DNA in a human being was actually being used.^5/

This juxtaposition of percentages suggests that the scientific community has shifted from the view that “only 5 to 10 percent” of the genome is functional (“needed” for the organism to function normally) to a sudden realization that 80% falls into this category.

But the more I read, the clearer it became that this description of a sudden phase transition in science is wildly inaccurate. Johns Hopkins biostatistian Steve Salzberg, in a penetrating and provocative Simply Statistics podcast interview, describes the 80% figure touted in the ENCODE paper as irresponsible.^6/ University of Toronto biochemist Lawrence Moran saw it a repeat of a similar, problematic performance five years ago, at the conclusion of the pilot phase of ENCODE.^7/ Responding to criticism, ENCODE Project leader Ewan Birney explained the new knowledge this way:

After all, 60% of the genome with the new detailed manually reviewed (GenCode) annotation is either exonic or intronic, and a number of our assays (such as PolyA- RNA, and H3K36me3/H3K79me2) are expected to mark all active transcription. So seeing an additional 20% over this expected 60% is not so surprising.^8/

“Not so surprising”? A whopping 60%—not a minor 5 or 10%—was already estimated to be “active”? What is going on here?

The answer lies in the definition of some key terms (like exons, introns, and transcription) and requires a rudimentary understanding of the fundamentals of gene expression and its regulation in human beings. This posting presents the essential terminology and concepts. Part II will apply them to explain what ENCODE’s “assign[ing] biochemical functions for 80% of the genome” means. Anyone who knows what RNA transcripts and transcription factors do can skip this first part (or can read it to let me know of my inaccuracies).

To avoid suspense, I shall lay out my conclusions here and now: (1) if ENCODE gives a clear number for a percentage of the genome that regulates genes—the promoters, enhancers, silencers, ncRNA “genes,” and so on—I have yet to find it; (2) this number is almost surely less than the 80% figure reported for functionality; and (3) “functional element” as defined by the ENCODE Project is not a term that has clear or direct implications for claims of the law enforcement community that the loci used in forensic identification are not coding and therefore not informative. Those claims of zero information are somewhat exaggerated, but that is another story. For now, I merely describe some basics of gene expression and regulation.

Genes make proteins. But how? There are three big steps (with many activities within each step): transcription; post-transcription modification and transportation; and translation. All involve RNA, a single-stranded molecule related to DNA, and proteins. The basic picture is

• Transcription to precursor messenger RNA: DNA + proteins --> pre-mRNA (in nucleus)
• Post-transcriptional modification and transportation: pre-mRNA + proteins and RNAs -> mature m-RNA (in cytoplasm)
• Translation to protein: mRNA + tRNA and proteins --> expressed protein (in cytoplasm)

In the first big step, the base pairs of the gene are transcribed jot-for-jot into an RNA molecule (precursor messenger RNA, or pre-mRNA). In the second major step, the transcript is modified at its ends, edited to remove parts that do not code for the protein that will be made (splicing), and the mature messenger RNA (m-RNA) is moved outside the nucleus. In the third phase, another type of RNA (transfer RNA, or tRNA) stitches together individual amino acids in the order dictated by the m-RNA transcript to form a protein, thereby translating the DNA sequence mirrored in the mRNA into the amino-acid order of the protein. Translation occurs on a kind of microscopic workbench (a ribosome) made of yet another RNA (ribosomal RNA, or rRNA).

For all this to happen, the DNA, which lies tightly coiled in the chromosomes (in a protein-DNA matrix known as chromatin), must open up for transcription to occur. Thus, changes in the chromatin regulate transcription, and these changes can be brought about in a number of ways. Transcription factors (specialized proteins) bind to the DNA. The bound transcription factors then recruit an enzyme (RNA polymerase) that produces RNA. This occurs within a region of DNA, known as a promoter, near the start of the protein-coding DNA (the structural gene). The level of transcription is influenced by activator or repressor proteins that bind to still other small regions (enhancers and silencers, respectively) that also lie outside the structural gene. In short, chemical interactions that open or close the chromatin that houses the DNA and transcription factors regulate the first step in the DNA-to-protein process.

In the past decade, other mechanisms of regulation or control of gene expression have been discovered. Many DNA sequences are not transcribed into messenger RNA, but they are transcribed into a variety of other RNAs. These non-protein-coding DNA sequences can be thought of as genes for RNA. Courting confusion, they usually are called “noncoding” (ncDNA)—because they do not code for protein—but they certainly code for RNAs that are crucial to translation—rRNA and tRNA—and for other RNAs that affect transcription, translation, and DNA replication. So it turns out that the genome is abuzz with transcription-to-RNA activity and other events that feed into the expression of the (protein-)coding DNA.

Yet, this hardly means that every biochemical event along the DNA is functionally important. Some, perhaps many, non-mRNA transcripts are just “noise.” They may float around for a while, but they may not do anything except wither away. In addition, large segments of the DNA transcribed in the course of making mRNA appear in the initial transcript (the pre-mRNA) but never make it into mature mRNA. These unused parts of the pre-mRNA transcripts correspond to long stretches of DNA, known as introns, that interrupt the smaller coding parts—the exons—that are translated into proteins. The initially transcribed intronic parts are removed from the pre-mRNA in a process called RNA splicing. Most of the RNA from introns probably just dissipates.^9/

All these terms are a mouthful, but armed with this basic understanding of genes, RNA, and proteins, we can see why the 80% figure does not mean what one might think. We shall also see that the estimated proportion of the genome that encodes the structure of proteins or regulates gene expression has not jumped from 5 or 10% to 80%.

Notes

1. Ian Dunham et al., An Integrated Encyclopedia of DNA Elements in the Human Genome, 489 Nature 57 (2012).

2. E.g., Elizabeth Pennisi, ENCODE Project Writes Eulogy for Junk DNA, 337 Science 1159 (2012).

3. E.g., Gina Kolata, Bits of Mystery DNA, Far From ‘Junk,’ Play Crucial Role, N.Y. Times, Sept. 5, 2012. In one respect, the "dark matter" metaphor misrepresents dark matter. The presence of dark matter is inferred from its gravitational effects on visible matter. The presence of noncoding DNA is known from experiments that detect and characterize it just as they do coding DNA. Perhaps the metaphor means that the sequence of “dark matter” DNA cannot be deduced from the structure of a protein made in a cell. This, however, is like saying that dark matter is matter than cannot be seen with the naked eye. And that is not what astronomers mean by dark matter.

4. E.g., House Committee on the Judiciary, Report on the DNA Analysis Backlog Elimination Act of 2000, 106th Cong., 2d Sess., H.R. Rep. No. 106-900(1), at 27 (“the genetic markers used for forensic DNA testing … show only the configuration of DNA at selected ‘junk sites’ which do not control or influence the expression of any trait.”); New York State Law Enforcement Council, Legislative Priorities 2012: DNA at Arrest, at 5, http://nyslec.org/pdfs/2012/1_DNA_2012.pdf (“The pieces of DNA that are analyzed for the databank were specifically chosen because they are ‘junk DNA.’).

5. Kolata, supra note 2.

6. Interview by Roger Peng with Steven Salzberg, podcast on Simply Statistics, Sept. 7, 2012, http://simplystatistics.org/post/31056769228/interview-with-steven-salzberg-about-the-encode (“Why do they feel a need to say that 80% of the genome is functional? … They know it’s not true. They shouldn’t say it. … You don’t distort the science to get into the headlines.”).

7. Laurence A. Moran, The ENCODE Data Dump and the Responsibility of Scientists, Sept. 6, 2012, http://sandwalk.blogspot.com/2012/09/the-encode-data-dump-and-responsibility_6.html (“This is, unfortunately, another case of a scientist acting irresponsibly by distorting the importance and the significance of the data.”).

8. Ewan Birney, ENCODE: My Own Thoughts, Sept. 5, 2011

9. Post-splicing processing of a small fraction of the RNA from introns can produce noncoding RNAs that may regulate protein expression. L. Fedorova1 & A. Fedorov, Puzzles of the Human Genome: Why Do We Need Our Introns?, 6 Current Genomics 589, 592 (2005).

I am grateful to Eileen Kane for explaining some of the molecular biology to me.

Trashing Junk DNA

You have seen the headlines:

• Bits of Mystery DNA, Far From 'Junk,' Play Crucial Role (New York Times)
• 'Junk DNA' Concept Debunked by New Analysis of Human Genome (Washington Post)
• 'Junk DNA' Debunked (Wall Street Journal)
• Breakthrough Study Overturns Theory of 'Junk DNA' in Genome (Guardian)

Or maybe you heard MSNBC report that the data from ENCODE "shows us living beyond our genes" --whatever that means -- or listened to CBC intone that "'Junk DNA has a purpose" -- sounds divine -- or saw the Independent's mishugina announcement that "Scientists Debunk 'Junk DNA' Theory to Reveal Vast Majority of Human Genes Perform a Vital Function!" -- like we did not know that genes were functional and important?

The level of hype here is phenomenal. (Some useful clarification can be found at the Nature News blog.) In the next few days, I hope to post some quick thoughts on what the ENCODE figures (like 80%) being bandied about for the "functional" or "biologically active" fraction of the human genome mean for the loci used in forensic DNA identification.

(If any readers have insights to share, post a comment or send me an email at kaye at alum.mit.edu, and I'll try to use them. I am still educating myself about some of the details of gene regulation and can use any help I can get.)