Consequences of Infidelity Spellchecking DNA Thomas Kunkel, NIH Scientist

Gtgt Well the most fun part of the job for sure is working at the NIH. It gives me the opportunity to try to understand at a very basic level how nature works. Right now there are experiments going on behind me that will teach us something that no other human being has ever known. We now combine structural biology, biochemistry, molecular genetics, and sometimes even studies in whole animals like mice to study the genes and the biochemical reactions that achieve very high replication fidelity. So one of the goals of the DNA replication fidelity group is.

To get very high resolution crystal structures of DNA polymerases as they're making new DNA. That can be very expensive and can often fail. And this piece of equipment greatly increases our chances of success and greatly decreases the cost of that effort. So what this robot is capable of doing is setting up a very large number of different conditions in an attempt to get a good enough crystal of a DNA polymerase to get the kind of information we need to understand why defects lead to things like cancer.

And other adverse health consequences. This is Marta assumed spelling who recently joined my lab from Warsaw, Poland and she is constructing a yeast strain that is disabled in a gene that we think is important for human health. The little colonies growing on these Petri dishes are individual colonies of yeast cells and the one in my left hand contains a very large number of colonies and that's because those cells are growing on a Petri dish containing everything the cell needs to be happy and grow well rich media.

The Petri dish in my right hand contains many fewer colonies and the reason that's so is because even though we put a very large number of yeast cells on that Petri dish, only a few of them can grow because they have a mutation in an important gene that allows them to grow on that kind of medium. So the difference between many colonies and few colonies can be measured, quantified, and used to calculate the actual mutation rate in this yeast strain resulting from inactivating a gene that's important.

Why Are We Addicted to Gasoline

Why are we addicted to gasoline There are many reasons to break our dependence on gasoline, so why do we keep using it Because it's liquid magic! It's amazing how much internal energy it has. A pound of gasoline has nearly ten times the energy of a pound of TNT. In automobiles, gasoline provides the power to turn the drive shaft. I can spin this thing about 60 revolutions per minute. And that's with very little torque on it. A car engine drive shaft has to turn much, much faster, with a MUCH higher load.

Think about it, a simple tank of gasoline can turn this car's drive shaft at 3,000 revolutions per minute.for hours on end. I'm not saying that gasoline engines are the best solution. We know they have harmful byproducts. Yet they continue to be common because of the amount of lowcost energy they create. Let's look at the energy density in a gallon of gas compared to a nickelmetal hydride battery found in many hybrid cars. The battery contains less than a third of a megajoule of energy per kilogram. Gasoline contains 44.4 megajoules per kilogram.

Gasoline packs 153 times more energy in the same mass. That's like the difference between lifting to the same height this 20 pound dumbbell versus this 3,000 pound car. Your garage door spring uses mechanical energy to raise a heavy garage door. Could we build a car that runs on garage door springs If we do the math, we can see that you would need over halfamillion pounds of springs to hold the same amount of energy in one gallon of gas. And since we have relied on gas to power our cars for over 100 years, we have an infrastructure.

Jessica Williams of NIEHS discusses DNA replication

I'm Jessica Williams, and I am a research fellow in the DNA Replication Fidelity Group, which is headed by Tom Kunkel. and so, what our lab works on is how DNA in a cell gets copied and, what happens is that if it doesn't get copied accurately, this can result in mistakes in the DNA, which we call mutations. and what we found in this study was that one reason why these mutations are created is due to the activity of a protein, which targets the specific type of error that's been introduced by the DNA polymerase.

One important health implication for these results is that there is a severe autoimmune disorder, AicardiGoutieres syndrome. And, so one possibility is that these patients have more of the types of mutations in their DNA that we identified in this study, so these short deletions in repetitive sequences. This work was a collaboration between our lab, Sue JinksRobertson's lab at Duke University, and Yves Pommier at the NIH. What we found was that when we made yeast strains that lacked the ability to remove a specific type of error in their DNA,.

We got mutations, and these mutations were gaps in DNA. And, what we identified in this paper was the protein that's responsible for generating these gaps, and it's called topoisomerase 1. In this model for topo 1initiated deletions, we have an AT, AT repeat sequence. And, normally, for example, the second A of that AT, AT, a T is incorporated opposite by the DNA polymerase. However, we know that in some cases DNA polymerases can incorporate a ribonucleotide, wherein in this model, it's a riboU. If this is a site for Top1 cleavage,.

It will bind and cleave however, in the example where you have the riboU present, a break in the DNA is created. End processing can then occur, and in the next step, a gap is now left in the DNA, where one of these AT repeats was located. The DNA strands can then realign And you see loss of one of these AT repeats And then finally, ligation and replication results in loss of one of the ATs of the repeat, and a twobase pair deletion. The results from this study raise a number of important questions to be addressed.

Building Microscopes that See into Tiny Brains Hari Shroff, NIH Scientist

Gtgt My name is Hari Shroff and I work in the Intramural Program at the NIH, in the National Institute for Biomedical Imaging and Bioengineering. And what my lab tries to do is to develop new forms of microscopy that let biologists and medical researchers at the NIH and around the world look at things that were too small or too fast or too delicate to be seen with normal kinds of microscopes. There are a couple of things that are really spectacular about working at the NIH. One of them is that my background is sort.

Of in biophysics and I'm, I would say, equally at home discussing biology and doing physics and engineering, but really the kind of bread and butter tools that we use are physical tools and engineering tools. And for somebody like me that tries to make better tools for biologists, the NIH is a great place because we're surrounded by biologists that are investigating all sorts of different systems. Everything from molecules to cells to whole animals, and so this is a great environment for trying to find problems and for trying to really get at, you know,.

What are the technology issues that have to be addressed so we can explore that next frontier of biological imaging. gtgt If you want that clicking to startup again. gtgt See all this stuff over here This is that thing where that weird, remember this gtgt Oh yeah! Biologists want to look at a wide range of samples. Sometimes you want to look at single cells, but sometimes you want to look at tissues or even whole organisms. The type of microscope we were trying to improve upon, up until now, has only been suitable for single cells.

So what we've done here is we've extended we preserved all the good qualities of that microscope, we hope. It's about as fast. It has about the same resolution. You see just about as well. But you can look much deeper. So instead of looking at a single, you know, human cancer cell line that's been preserved, we're looking at a worm embryo developing, you know, from a single cell up to a hatched adult, or a hatched adolescent. Or we're looking at a zebrafish embryo and an animal with organs.

And blood cells and an eye and a brain. gtgt The other sort of more practical aspect that is really great about being part of the Intramural Program is the sort of freedom to actually devote 90 percent of my time to actually doing research. One of the reasons that I came here instead of going to like a more traditional university and becoming an assistant professor is that here I'm in a way spoiled. I really don't have to write grants or teach, and I can devote sort of almost all my time.

The Sperm Struggle is Real

Sperms aren't the mindless, flagellumwielding, dolts you thought they were. They are wily little devils with war on their nuclei. Music Hey dudes, Trace here with the lowdown on your sperms' struggles for DNews. Sperm are the male gamete, the counterpart to the female's egg. When a sperm reaches the inside of an egg, a zygote is created the first step on the path to a fetus. But to reach that goal, a sperm have to overcome a lot of different problems one being there are millions of other sperm. They're like, WATCH OUT DUDE. Imma tryin' to get to the egg!.

For instance, we've evolved what science calls sperm allocation. As the point of sex is to get your sperm to the egg first and thus spread your genes, sperm allocation was one of nature's ways of getting the most bang for your. buck. IF a male believes his female sexual partner has committed infidelity, or if he's simply spent a long while away from the partner, his sperm count will increase when they pair up! The authors of a study published in Current Directions in Psychological Science believe this is why males get lustful feelings after.

Being away from their partner they want to make sure they can get as much sperm in there as possible to compete. Once inside the woman, sperm continue the struggle. This has to do with that creation sperm are formed based on HOW the species fertilize AND how much sexual competition is present. It was assumed that the bigger the sperm, the faster they are. Bigger and stronger is better Right Wrong. A study published in the journal Evolution uses Bonobos as an example. These apes share almost 99percent of their DNA with humans and have a lot of.

Sex with lots of partners. The competition has evolved sperm with bigger head and a smaller tails. The larger sperm were actually SLOWER. The researchers found only animals who fertilize OUTSIDE of their bodies have a large sperm advantage. The shorter head and longer tail of fish and squid sperm were super fast and thus got to the egg first. Yah bro! Then there's the theory that there are two kinds of sperm the goal scorers, and the ones who play defense. Of the 250 million sperm cells released into the female reproductive.

System, 27percent are defective, right off the bat. The normally ovalshaped head is spherical or quite pointy in parts, the tails are malformed or whatever. This theory of so called kamikaze sperm also comes back to competition between mates. The idea is that these malformed sperm hang back to block those from competing mates, thus ensuring the first male's DOMINATION OF FERTILIZATION!!! Since the hypothesis was proposed in the 80s, scientists have tried and failed to reproduce their findings, so while we're pretty sure this isn't real, it's still being tested.

Most scientists now believe the 27percent defect rate occurs because of poor quality control in sperm creation. Which. is a little embarrassing. Dudes, this is our big moment and we're halfassing it out there! 110percent, c'mon! Of the millions of sperms released, remember you were the fastest. So tell us your strategies in the big empty white box below and subscribe! Also, double check you're still following us on Twitter AtDNews. We made some changes. You can get behind the scenes pictures and cool stuff that never makes it into an episode. Thanks!.

Cellular Stress, RNA Metabolism and Aging Myriam Gorospe, NIH Scientist

Gtgt Where is a given messenger RNA Is it in the cytoplasm or the nucleus How much of it is there And in the context of aging, we are interested in understanding how our cells respond to stress. By comparing how a young cell responds with how an old cell responds, we're getting very useful information about what it is that we lose as we age. What are the types of responses that our body is no longer able to mount properly as we grow older So the stress that we have used to generate the image.

That you see here is arsenide. Arsenide is a very inaudible oxidant and oxidated stress is one of the stressors our bodies have to cope with during our lifetime. So when we treat these cells with arsenide, something very interesting happens and that is that the RNA in the cytoplasm goes to this distinct fosite assumed spelling set of inaudible. Fosite.and we know it's a cytoplasm because it's outside of these round larger circles that represent the nuclide so now the RNA is assembled in these fosite that we call stress granules..

And for this image, we used antibodies that recognize two proteins called A2 and TA1 assumed spelling that we know based on past work and work of other labs as well as our own, we know that A2 and TA1 both distress granules and they are.they retain the messenger RNA until the stress in the cell has resolved and the cell has fixed the damage that took place because of the arsenide treatment and these structures that look very nice and distinct will actually dissolve and disappear. So if we came back two hours later, we would see none.

Of these stress granules they will be all dissolved into the cytoplasm. I think that NIA is very unique in that you have.for example the NIA is a good example you have an institute where people with very different skills, very different approaches to science, very different study model systems, they all have in common the same interest in understanding the process of aging. So we have expertise in cellar molecular biology, but we can collaborate with colleagues down the hall who work with animals and then we can sort of ask the same questions in an animal model.

Thomas Kunkel Explains Deoxyribonucleic acid DNA NIH Scientist

Gtgt So in this structure Kasha assumed spelling was showing you, all this blue is DNA. DNA is deoxyribonucleic acid. So the phrase deoxy comes from the fact that on this sugar which is a component of the sugar phosphate backbone of DNA, there's not an oxygen on that position and oxygen is depicted in this image as red. So there's no oxygen on this position which is called the two prime position, but this polymerase is busy putting in a ribonucleotide which in DNA is a mistake because at that same two prime position there is an oxygen.

And this is typical of RNA but not DNA. That's bad news for a cell potentially because chemistry teaches us that if there's an oxygen on this position, it can lead to breakage of the DNA and so organisms that have DNA as the storage medium for genetic information try to avoid putting ribonucleotides into the DNA because that would potentially break the DNA and lead to either mutations or cell death. So what we're trying to understand is how DNA polymerases incorporate or do not incorporate the incorrect sugar into DNA.

And that has.that subject started four years ago with the work of a summer student, Mike Andrew, working with a postdoc in the lab who discovered to our amazement that the DNA polymerases that are busy replicating the nuclear genome in higher organisms like humans put a lot of ribonucleotides in the DNA. That was a big surprise. And so what we've done since that summer of 2009 is spent a very large amount of time by the people around the table here trying to understand how the ribonucleotides get in,.

How many of them get into DNA, where they are in DNA, and how they're removed from DNA, and lastly if they're not removed, what kind of either good or bad consequences might that have for a cell, including a human. And a reason why that is very, very interesting in terms of the biomedical community is that we know that the enzymes that.if this is.ribo is accidently put into the DNA, the humans have enzymes that remove it. And it was discovered as far back as 2006 that the enzyme.

Divorce Advice Top Reasons for Divorce

The top reasons for not winning the lottery are because the odds are against you. The top reasons for divorce well that's another story. And this is Dr. Paul author of Boomer girls, a boomer woman's guide to men and dating and host of ask Dr. Paul. The top reasons for divorce, somehow I feel like one of the comedians who comes up with the top ten reasons. And we won't go there on a comic way because it's not funny.The number one is adultery. Number one is cheating. Ok. That's number one. We are not going to go through top ten.

But childcare is another one. They disagree on how to raise children but that's way down the list. Finances is another one. Ok. That's probably number two in that top ten if you will and we are not going to go through ten but number two is having a real financial difficulty with each other. One is a saver and one is a spendthrift and it doesn't work. Or you've gotten yourself so deeply into debt that you are arguing about it all the time. It's a bad, bad scenario. Number three oddly enough, number three comes under the category.

Of addiction. And it can be an addiction that maybe minor, one person sees it as minor, the other sees it as major. Be it alcohol, or drugs or gambling or smoking. The smokernonsmoker is a recipe for divorce beyond belief. Another issue one of the top issues in getting divorced is risk taking. Suddenly one person wants to go into business and is willing to take a big risk and mortgages the farm, the house whatever and the other one says, whoa stop right now I'm not a risk taker. Those things should have been probably exercised in terms.

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