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James A. Shapiro

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Epigenetics I: Turning a DNA Packaging Problem Into a Developmental Control System

Posted: 07/09/2012 6:38 pm

Two postings back, I promised a commenter called Sierkovitz that I would discuss epigenetics. This is an important subject with major implications for understanding natural genetic engineering in evolution. So here is the first of at least three related blogs.

"Epigenetics" literally means "over or above genetics." It refers to hereditary changes in genome expression that do not involve alteration of DNA sequences.

Contemporary ideas about epigenetics have two independent historical sources that have subsequently merged in a remarkably satisfying way. The first source was theorizing about cell differentiation and morphogenesis by Conrad "Hal" Waddington, one of the most imaginative and penetrating mid-20th-century geneticists. Waddington realized that a heritable control process was necessary for cells with the same genome to form tissues containing different kinds of cells. In 1942 he called this the "epigenotype," meaning a higher-level regime placed over the genome during development so that different sequences could be expressed in distinct cell types.

The second source of epigenetic ideas came from observations on DNA packaging in the cell. The DNA in our cells would be over 6 feet in length if stretched out, but the nucleus is only about 1 ten-thousandth of an inch across. Clearly, our genomes are densely compacted to fit in such a small volume. Moreover, the packing has to be highly organized so that replication, transcription, chromosome movements, and all other genome functions proceed smoothly.

The historical reality is that cytogeneticists (literally, cell geneticists) had been observing DNA compaction since the 19th century through their microscopes. They described various forms of "chromatin" (i.e., colored material) along the length of chromosomes. The prefix "chroma-" refers to the coloration of chromosomes by various stains used to make them visible. Normal staining was called "euchromatin" (i.e., "true" chromatin), and darker staining was called "heterochromatin" (i.e., "different" chromatin).

Using distinguishable chromatin regions in her maize stocks, the pioneer cytogeneticist Barbara McClintock and her student Harriet Creighton were the first to demonstrate that chromosome physical structure corresponds to a genetic linkage map. From studying what was initially considered a marginal phenomenon in genetics, "position effect variegation," geneticists came to understand that differences between eu- and heterochromatin had a profound impact on genome expression.

Today, we understand that the molecular basis of DNA compaction into chromatin provides the epigenetic control system that Waddington first postulated in the 1940s. The way the chromatin forms regulates how accessible the chromosomal DNA is to proteins and RNA molecules that carry out replication, transcription, repair, recombination, natural genetic engineering, and attachment of protein motors and filaments for moving the genome within the nucleus.

During cell differentiation and development, distinct cell types "index" different regions of the genome into expressed and unexpressed chromatin domains. Thus, the set of encoded functions can be "canalised" (Waddington's term, with British spelling) into those appropriate for each specialized cell type. There are special signals and processes that punctuate the genome for formation into chromatin domains that may span a significant number of separate coding regions.

DNA in chromatin is modified chemically and compacted in two ways:

  1. By having methyl (-CH3) groups added to or removed from one of the bases (DNA methylation and demethylation), and
  2. By being tightly wound into "nucleosomes" (literally "nuclear bodies") around specialized positively charged proteins called histones. The negative charge on the DNA attracts it to the positive charges of the histones. The nucleosomes are further grouped into several levels of higher-order folded structures that help achieve the tremendous compaction required to fit the DNA into the nucleus.

Cells control chromatin structure exquisitely. They have a chromatin formatting and reformatting system that is a wonder of molecular signaling and control. There are arrays of specialized "chromatin-formatting" enzymes that add or remove methyl groups from the DNA and other enzymes that add or remove various chemical groups from specific amino acids in the "tails" of the histones that peak out from the nucleosomes. These covalent (stable) chemical modifications of the DNA and the histones constitute an intricate code that the cell can read to determine the accessibility status of the underlying DNA, independently of its sequence.

Chromatin structures can be inherited or changed during the life cycle and across generations. This is most easily appreciated from the phenomenon known as "imprinting." The expression of many genetic loci does not depend on the DNA sequence alone; it also depends on which parent contributed it to your genome. When sperm and egg cells form in male and female gonads, particular regions of the genome are epigenetically imprinted into special chromatin domains so that they will either be expressed or not post-fertilization, during embryonic development and after birth. Some loci are only expressed from the father, and some only from the mother. The imprintings are stably inherited through all the cell generations it takes to make an individual.

We are beginning to discover how many epigenetic configurations are inherited across generations. This is the kind of inheritance that can have a profound influence on evolution, because it can be selected and expand in a population. Some very stable multigeneration genome changes are, in fact, "epimutations" that do not involve any DNA sequence alteration, only chromatin reformatting.

Other transgenerational changes are reported to be induced by a wide range of environmental factors, such as environmental stress, endocrine disruptor chemicals, infection, and even maternal grooming. The ability of environmental factors to cause inherited epigenetic modifications makes this form of genome coding particularly attractive to neo-Lamarckists.

There is a great deal more to say about epigenetic inheritance. In the next couple of blogs, I will discuss epigenetic memory and targeting, overturning a six-decade mainstay of evolutionary dogma, and the connection between epigenetic changes and natural genetic engineering of DNA sequence organization. Stay tuned for some eye-opening research.

 
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Two postings back, I promised a commenter called Sierkovitz that I would discuss epigenetics. This is an important subject with major implications for understanding natural genetic engineering in evol...
Two postings back, I promised a commenter called Sierkovitz that I would discuss epigenetics. This is an important subject with major implications for understanding natural genetic engineering in evol...
 
 
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Wendell Read
09:42 PM on 07/12/2012
James,

This article points out an interesting parallel between epigenetic factors controlling cellular development and memory/cognitive functions.

http://www.cell.com/neuron/retrieve/pii/S0896627311004338#MainText

An example:

"Investigation of the precise molecular mechanisms in both cellular development and memory has increased over the past two decades, and an interesting new understanding has emerged: developmental regulation of cell division and cell terminal differentiation involves many of the same molecular signaling cascades that are employed in learning and memory storage. Therefore, cellular development and cognitive memory processes are not just analogous but are homologous at the molecular level."

The authors are of course not talking about natural genetic engineering (NGE), but could some of their findings apply to NGE?
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James A. Shapiro
Author "Evolution: A View from the 21st Century
12:31 AM on 07/13/2012
Wendell,

I'll discuss at least some of what we know about epigenetic control of NGE in the third blog of this series. Maybe you'll see a relationship there, but I think more work needs to be done first like the exploratory experiments outlined in the last two blogs. Once we have evidence for coordinated changes in real time, it becomes possible to investigate what brings the coordination about.
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bertvan
http://30145.myauthorsite.com/
01:50 PM on 07/12/2012
This is off topic, and begging your indulgence again, but you are one of the few biologists I trust. I seem to remember hearing that meiosis occurs twice in the female egg, once very early (at conception?) and the second time just before ovulation. Is that true?
Berthajane Vandegrift
A Few Autistic Questions about Freud, Marx, and Darwin.
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James A. Shapiro
Author "Evolution: A View from the 21st Century
02:26 PM on 07/12/2012
Berthajane,

Meiosis reduces the number of chromosomes in each cell by half and so produces gametes that are called "haploid." The reduction occurs because meiosis involves only one round of DNA replication that doubles the genome but two separate cell divisions that reduce the number of chromosomes by a factor of four. The two divisions thus end up with a net halving the number of chromosomes in each cell.

Although I am not an expert and may be wrong, I suspect what you heard is that meiosis in females is halted after the first meiotic cell division early in life. There is then a long period, and the second meiotic division occurs just before the haploid eggs are released into the uterus at ovulation for fertilization.
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bertvan
http://30145.myauthorsite.com/
03:29 PM on 07/12/2012
Don't you wonder if that second meiosis might be an opportunity to reorganize the genome while integrating some of effects of the mother's life experiences up to that time? If one believes life is a rational, purposeful process, surely there must be some reason for the second meiosis.
Berthajane Vandegrift
03:50 PM on 07/11/2012
James, have you seen this?

New work from Howard Hughes Medical Institute (HHMI) scientists suggests those abundant molecules (piRNAs) may be part of the cell's search engine, capable of querying the entire history of a cell's genetic past.

“This is really remarkable. It implies that an organism has a memory of all the previous gene sequences it’s ever expressed before.”
Craig C. Mello

http://www.hhmi.org/news/mello20120625.html

wow!

harry
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James A. Shapiro
Author "Evolution: A View from the 21st Century
09:12 PM on 07/11/2012
Harry,

Welcome back and thanks for the link.

The next blog deals with just this kind of epigenetic memory. The emphasis is more on DNA sequences that have come in from outside, but the process is basically the same.

Your "wow!" validates my promise, "Stay tuned for some eye-opening research."
06:47 AM on 07/12/2012
james,

thanks. I'll definitively stay tuned.
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SolarPowerGuy
Ph.D., Immunology; Solar power @ home; Green Party
07:11 AM on 07/13/2012
Wow, I wasn't aware of that.

If humans also have a piRNA system, the implications for cancer are ominous. As I recall, quite a few tumors are "reverted" cells, with properties which resemble embryonic stem cells. The piRNA system could let such cells sail right through, since the embryonic genes had been expressed once before in an ancestral cell.
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James A. Shapiro
Author "Evolution: A View from the 21st Century
05:40 PM on 07/13/2012
Solar,

As I replied to Harry, the piRNA system is a kind of immune memory of past invaders, not of expressed functions. In the few cases I know about, the piRNAs serve to direct silencing, not permit expression. They are also specific to the germline and would not be active in somatic tissue that becomes cancerous.

So, please don't lose any sleep over this part of epigenetic memory. We do know there are profound epigenetic changes in tumors, and there may be occasion to visit them in the future. But I need to learn more about the subject myself.
09:19 AM on 07/11/2012
Dr. Shapiro,

I look at the human body, where each cell has 750MB of code and each different type of tissue is built by epigenetically expressing different portions of that code. I see an amazing efficiency. The genome is 750MB and Microsoft Windows is 25 gigabytes; humans are certainly superior to Windows.

As a person who builds all sorts of systems, trying to make use of every resource I have, it makes me wonder: Trillions of cells, each with the same 750MB of code, seems excessively redundant.

However if each one of those cells could epigenetically apply a unique context to its own information in real time, thus storing more information, you have an incredibly clever mechanism for archiving not just terabytes but petabytes of additional data, epigenetically distributed across billions of cells.

And if those same cells can communicate with each other, it might better explain how an entire human body adapts to, say, training for a marathon. The stress affects every cell in the body, each in a slightly different way. Thus an entire organism would have more than enough space to store incredibly detailed information about its own unique context in the world.

Then perhaps the very most important adaptations would be passed to offspring, IF the signalling systems could coordinate appropriate information to the reproductive cells.

I suppose all this could sound far-fetched to some. But is there evidence that any of this could be true?
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James A. Shapiro
Author "Evolution: A View from the 21st Century
11:00 AM on 07/11/2012
Perry,

I was glad to see that you pointed out how the epigenetic encoding tremendously increases the capacity and flexibility of the genome as a RW storage system. With all the possible chemical modifications of the histones, the combinatorial possibilities are immense, and there are also specialized histones for certain tasks, like DNA repair.

As for passing on the effects of life history experiences to future generations, see the links provided by Rhynocstylus below. Amazing examples in a long-term study of an isolated Swedish population. Now that we have mechanisms for non-DNA inheritance, we will see many more well-documented cases emerging in the literature. Some of the laboratory studies are also posted at http://shapiro.bsd.uchicago.edu/ExtraRefs.GenomeCompactionChromatinFormattingEpigeneticRegultion.shtml under the heading "Epigenetic responses to stimuli, epigenetic memory, and transgenerational effects."
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khamilton0231
Can't we all just get along?
08:35 PM on 07/10/2012
A simple yet delightful introduction into the multi-layered "onion" that is molecular biology.
06:02 PM on 07/10/2012
Prof. Shapiro,

Re: "Cells control chromatin structure exquisitely. They have a chromatin formatting and reformatting system that is a wonder of molecular signaling and control."

Formatting, signaling and control would seem to be the result of intelligence and forethought. what is the evolutionary explanation for those attributes?
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Gas-Bag
There's nothing endearing about perfection.
10:54 PM on 07/10/2012
That's the bit that gets everyone arguing around here, I think that the good Professor thoroughly enjoys it :-)
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James A. Shapiro
Author "Evolution: A View from the 21st Century
11:55 PM on 07/10/2012
Gas,

If you'll forgive a pun, I find it a gas when people want to learn about the nitty-gritty of what cells can do with their genomes. Gives a whole new meaning to the idea of what life is all about.
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Wendell Read
01:42 AM on 07/11/2012
It is of course a fascinating question: We have two choices, our metaphysically based 'ground rules' can constrain how we interpret new data - certain 'explanations' of the data are ruled out even though they would otherwise be the most obvious way to understand them, or we can set aside all 'ground rules' and investigate the clear implications of the new data that are being collected.

Does the good Professor thoroughly the fallout from this basic question? I am sure he does. Science advances by the willingness of investigators to proceed in the face of 'received dogma' which constrains the interpretation of new data to the strictures of old and failing paradigms.
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James A. Shapiro
Author "Evolution: A View from the 21st Century
11:52 PM on 07/10/2012
Bill,

We still lack a detailed explanation for the origins of this remarkable control system. We can see simpler versions of it in yeasts and other microbial eukaryotes and can observe how the system becomes more complex through events like duplications and domain shuffling of the proteins involved.

Here are some links to recent papers that may give you some leads if you can navigate the PubMed database. I don't have space to provide the complete references: http://www.ncbi.nlm.nih.gov/pubmed/21507350, http://www.ncbi.nlm.nih.gov/pubmed/20097869, http://www.ncbi.nlm.nih.gov/pubmed/20210320, http://www.ncbi.nlm.nih.gov/pubmed/21119628, http://www.ncbi.nlm.nih.gov/pubmed/21596317, http://www.ncbi.nlm.nih.gov/pubmed/21647299, http://www.ncbi.nlm.nih.gov/pubmed/22388813, http://www.ncbi.nlm.nih.gov/pubmed/21663790, http://www.ncbi.nlm.nih.gov/pubmed/20738881, http://www.ncbi.nlm.nih.gov/pubmed/21119629, http://www.ncbi.nlm.nih.gov/pubmed/21527910.

As you might imagine, the literature on this subject is vast, and the field has not yet been able to digest if all from an evolutionary perspective.
09:40 AM on 07/11/2012
Prof. Shapiro,

Thank you for your comment and for the hyperlinks. That should provide enough research material to keep me off the streets for some period of time.
09:16 AM on 07/10/2012
Nice to see my (nick)name in it:)

But I think my original question might have been misunderstood to some extent. My thoughts were rather going along the lines - if the mechanisms responsible for the epigenetic mechanisms are themselves encoded - can they still account as if they "do not involve any DNA sequence alteration, only chromatin reformatting.".

Or is it also plausible that the genetic changes involved in conservation of epimutations are just not as straightforwardly linked to them as the regular mutations (eg. epimutations are caused by regular mutations elsewhere, epimutations are a result of some changes in promoter sequences of regulatory genes etc.). The topic is quite interesting and somewhat linked to my research, at least conceptually.I research evolution of bacteria and how acquisition of new "allele" or variants in a population can happen via alternative routes rather than "regular" mutations. And the thought that continuously bothers me is - if let's say natural transformation is still a mechanism that is coded, if its prevalence can be traced to particular conditions and particular mutations - is it still horizontal or rather a laterally transferred but horizontally active system. Sounds bit confusing, but its a concept that I still have not tackled completely, so please do feel free to ask for clarifications where needed:)
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James A. Shapiro
Author "Evolution: A View from the 21st Century
01:26 PM on 07/10/2012
Sierkovitz,

If you follow the link on "epimutations," you will find a paper on "paramutation" by Vicki Chandler. This is one case where we know that the stable changes are in chromatin formatting and are not the consequences of sequence changes elsewhere in the genome.

It remains, true, however, that mutations disrupting the epigenetic silencing system can reverse a wide variety of epimutations (where the epimutant state is silent) and induce yet others (where the epimutant state is expressed). In the next blog, I will explain a case where a stable mutant state can be induced by disrupting epigenetic silencing and then reversed by targeted epigenetic silencing.

There are no clear examples of similar stable epigenetic changes in bacteria, but we do know about many loci where DNA methylation plays a key role in regulation and switching expression of particular sequences on or off (e.g. http://www.ncbi.nlm.nih.gov/pubmed?term=17220888).

Please say something about your research and how it may be linked to epigenetic regulation.
11:55 PM on 07/09/2012
Offspring of reef fish pre-adapted to CO2 changes:
http://www.sciencenews.org/view/generic/id/341931/title/Climate_adaptation_may_be_a_family_affair

Instant epigenetics!
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HUFFPOST BLOGGER
James A. Shapiro
Author "Evolution: A View from the 21st Century
01:34 PM on 07/10/2012
Brian,

Thanks for the link. This may be another case of transgenerational inheritance of epigenetic modifications. But that is not the only possible explanation. We are also beginning to find reports about transmission of small RNA molecules to offspring, and there may be yet other forms of non-DNA inheritance at work.

It is exciting that the molecular studies are beginning to give us ways to understand phenomena that we were taught for so long were unthinkable or had been disproven. My next blog will contain a different example of a previously "disproven" way that cells change their genomes in response to life history events.
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Wendell Read
10:57 PM on 07/09/2012
Sometimes we miss the import of certain biological facts. Dr. Shapiro points out that the the length of the human DNA molecule is about 6 feet. Our initial reaction might be "gee, that's nice to know". But, let's look a little deeper. The diameter of the nucleus (which contains the DNA) is about 6 microns. Lets multiply this by 10,000. We now have a diameter about 2 1/2 inches, about the size of a tennis ball. The DNA molecule, its length increased by the same factor, is about 12 miles. Now imagine 12 miles of very thin thread stuffed into a tennis ball. You are challenged to locate a small specific segment say startling at 6 miles, 1248 feet, 3.1473 inches from one end. Lots of luck in finding it in this enormous tangle. Yet, this is what is happening all the time in our cells. The way the DNA is first wound on histones and then this complex further coiled up to enable specific sites to be located is amazing indeed. What at first seems commonplace often turns out to be a marvel of engineering.
07:52 AM on 07/11/2012
Mr. Read, that is quotable. I think you should be a popular science writer :^>
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bertvan
http://30145.myauthorsite.com/
10:18 PM on 07/09/2012
The biological reproduction is complex, and most of us are not qualified to understand the details. It surely takes a lifetime of study. Today there are basically two competing concepts of reality. (1) The materialist view that reality is an impersonal mechanical contraption, without plan, purpose or meaning. According to this view, the universe can only change by pointless accidents popping into existence, and thus life, the most complex aspect of our universe, is driven by “random mutation and natural selection.” (2) The non materialist view is that intelligence and purpose are essential aspects of reality. Some religious people are content to assign all purpose to their god. Some of us religious agnostics see a designing intelligence as an innate aspect of living systems Life designs itself, intelligently and purposefully. For many years the atheistic materialists have insisted that “random mutation and natural selection” has been scientifically proven as the force driving evolution. I never believed that proof existed. Even though I still don’t understand the details, I’m thrilled to see Dr. Shapiro exploring mechanisms that seem compatible with the view that there is more to biological adaptation than RM&NS. The materialists are still quite entitled to their mechanical view of reality, but they are not entitled to impose it upon the rest of us as scientific fact.
Berthajane Vandegrift
A Few Autistic Questions about Freud, Marx and Darwin
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Rynchostylus
10:03 PM on 07/09/2012
I hope the author discusses the study of the isolated Swedish population re: transgenerational epigenetic "memory". Incredible work...
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James A. Shapiro
Author "Evolution: A View from the 21st Century
01:39 PM on 07/10/2012
Rhynco,

I am not familiar with the work you are mentioning. Please give us a reference, or even better a link. That way we can all see what you are bringing to our attention. Thanks in advance.
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Rynchostylus
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HUFFPOST COMMUNITY MODERATOR
M4dwoman
There's a hole in the bottom of the sea
08:37 PM on 07/09/2012
Well color me pleasantly annoyed.
I actually had to read the whole article and pay attention while doing it.
Can't wait to read the follow up articles.
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James A. Shapiro
Author "Evolution: A View from the 21st Century
01:42 PM on 07/10/2012
Woman,

You have warmed my heart. One of the reasons I started blogging was to learn how to communicate contemporary science in a way that is accessible to non-specialists. It's gratifying to hear that I may actually have succeeded. Thank you.