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  • Hi. It's Mr. Andersen and in this podcast I'm going to talk about DNA replication.

  • That's the process by which DNA makes a copy of itself. Why is that important? Well this

  • right here is an egg being fertilized by a sperm. That means it's about to become a zygote.

  • It'll divide through mitosis to eventually create an embryo, a fetus and eventually create

  • a human. And that human is going to have billions and trillions of cells. And we have to make

  • sure that each of those cells has the same exact DNA that was in that original cell.

  • And we do that through the process of DNA replication. If we were to point specifically

  • where that occurs, well in eukaryotic cells, like this little baby here, that cell cycle

  • basically is remember, the G1 phase where it grows. The S phase where we copy all of

  • the DNA. The G2 phase is where it continues to grow. So this whole process right here

  • is called interphase. We then have mitosis where we go through prophase, metaphase, anaphase,

  • telephase, cytokinesis. But right here we have to make sure during that S phase that

  • we copy all of the DNA. Now mitosis is not found in prokaryotes. But they're going to

  • use a process called binary fission. And if you look right here, here's their nucleoid

  • region. They'll copy their DNA perfectly before they split in half. And so in all life on

  • our planet, DNA replication is super important. And so basically when they figured out the

  • structure of DNA, three theories came about as to how it actually makes copies of itself.

  • The first is semi-conservative, conservative and dispersive. Watson and Crick actually

  • believed in this. They believed that DNA would split in half. And then you'd copy new strands

  • on either side. But there were other scientists who believed in a conservative theory that

  • that first DNA remains intact and it kind of makes a photocopy of itself. And then some

  • believe that there was kind of a combination of conservative and semi-conservative. That

  • chunks of it were being split between the two. And this had to do with, they thought,

  • the histone proteins and how the DNA wrapped around it. And so basically the whole thing

  • was figured out through the Meselson-Stahl experiment. Basically what they used was two

  • different types of nitrogen. Good old run of the mill nitrogen 14. And then nitrogen

  • 15. An isotope that's heavier than nitrogen 14. So basically they bred a bunch of e.coli

  • on nitrogen 15 until all of their DNA was nitrogen 15. They then put them on a broth

  • of nitrogen 14. And basically in that first generation if it would have been conservative,

  • we would have had one band that would have been totally heavy at this N15 line. And then

  • one that was at N14. But instead we got this intermediary amount of DNA. In other words

  • it was a mix of the two. And then through generation after generation after generation,

  • they were able to figure out that this is how DNA copies itself. It copies itself semiconservatively.

  • And so to look at that in a little better detail, basically here is your DNA. It's a

  • double helix. It will unzip in the middle. So you can see it's unwinding right here.

  • And then we're going to add new strands on either side. So we eventually start, excuse

  • me, we start with one strand and we're going to end up with two strands. Each of these

  • strands are identical to that first strand. And each of them are going to contain half

  • of that original DNA. So again it's semiconservative in nature. Before we actually talk about the

  • process of DNA replication, we should talk about DNA and the parts of DNA. Remember DNA

  • is going to have three parts to it. Your basically going to have a sugar. That would be this

  • deoxyribose sugar right here. I could circle it. This is going to be our deoxyribose sugar.

  • You're going to have a nitrogenous base attached to that. And then you're going to have a phosphate

  • group. And so there are three parts to every nucleotide. Again we've got a sugar and a

  • phosphate and then a nitrogenous base. But you could see that there's going to be another

  • nucleotide right here. And another nucleotide going to be found right here. So let me clean

  • that up a little bit. Because it's anti parallel in nature. In other words DNA runs next to

  • each other. So it's parallel. But it's antiparallel in nature. In other words the two strands

  • of DNA are actually running in opposite direction. And when I mean running, I mean chemically

  • running in either direction. So basically how do we tell which way it is going? Well

  • we do that based on the sugar. And so if we look at this sugar right here, this sugar

  • is going to have a carbon here. So we call that carbon the 1 prime carbon. It's going

  • to have a carbon right here. And we call that the 2 prime carbon. It's going to have a carbon

  • right there. We call that the 3 prime carbon. It's going to have a carbon right there called

  • the 4 prime. And then it's going to have a carbon right here. And that's called the 5

  • prime. And so basically if you look here that whole thing is going to run, from the let's

  • look way down here, 1, 2, 3 prime end. Oops. Let me go back. So that's going to run from

  • the 1, 2, 3 prime end right here all the way up to the 5 prime end over here. Because here's

  • that 5 prime. If we look on the other side of the DNA you can see it's running the opposite

  • direction. From the 3 prime to the 5 prime. And that's going to be important when we look

  • at DNA and how it copies itself. And so let's look at that. If we look on the next slide,

  • in DNA replication there are tons and tons and tons of enzymes that are helping out.

  • It's way more complex than this. But from a diagram level this is pretty good. So basically

  • the DNA is going to be a double helix in this direction. But were going to have this enzyme

  • right here. It's called helicase. And it's basically going to unwind the DNA. So we're

  • going to go from this double helix to these single strands of DNA on either side. These

  • strands are going to be held in place using these enzymes. They're called single strand

  • binding proteins. They're basically going to hold it in place. And so now we have that

  • unwound DNA. The big enzyme that's super important in here is called DNA polymerase. So if we

  • look on this side, DNA polymerase is going to race down the DNA and it's going to add

  • new nucleotides on the other side of the DNA. So here's the original strand. And you can

  • see that DNA polymerase has already been here because it's added new strands in this direction.

  • Now the trick is that we can only add new nucleotides on the 3 prime end. We can't add

  • it on the 5 prime end. And so basically again. So here's the 5 prime end. If we follow that

  • right down here we can add DNA on this side, on the 3 prime end and it's just going to

  • go on silk smooth. In other words helicase is unwinds it. DNA polymerase adds the new

  • letters. And on this side we call that the leading strand. Everything is going to be

  • perfect. It's just going to flow on there perfectly. But the problem is since we can

  • only add DNA on the 3 prime end, we can't add it up here. We can't add it on the 5'

  • end over here. And so what's evolved is this really elegant method called the lagging strand.

  • So we can finish out the other side. And it's lagging strand because it tends to lag behind

  • the other side. If you have done any sewing, which I never have, it's kind of like back

  • stitching. In other words you're going in this direction but you're back stitching the

  • way as you go. And so basically there are a number of different parts that are found

  • in here. First thing that we have to do is we have to put down a primer. And so there's

  • going to be DNA or excuse me, RNA primase. And primase is going to add down a primer.

  • A primer is just one little bit of RNA. So we'll add a little bit of an RNA first. And

  • after we've added that RNA primer, then DNA polymerase can go in this direction. So once

  • the primer is in place, then we can run in that direction. And we can run in that direction.

  • We can keep running in that direction. So we've got to put a little RNA down and then

  • DNA polymerase goes. Unfortunately it can't connect it here. Because we've got DNA bumping

  • into RNA. And so there's going to be another enzyme. And that enzyme is called, let me

  • find it, DNA ligase. And so basically what DNA ligase is going to do is it's going to

  • go after that and clean up all of these messy junctions here. And it's going to put DNA

  • straight across it. And so basically that's a lot of stuff going on. What is all of that

  • doing? It's making sure that that message that was found in the DNA is copied to that

  • two new strands of DNA on either side. And there's some videos out on YouTube about how

  • DNA replication works. And they put together some computer animations of it, and it's wild.

  • It doesn't look like this at all. You have the lagging strand coming back upon itself.

  • So it's pretty amazing. Or you could even read the story of Okazaki, the person who

  • came up with this idea of how these Okazaki fragments work. Another fascinating story.

  • But we've got to finish. So basically what I want to talk about is origins of replication

  • or where DNA replication starts. Well in life there are basically two life types. We've

  • got the prokaryotics, which is going to be the bacteria and the archaea. And then eukaryotics

  • and that's going to be like you. And if you're prokaryotic you're going to have a single

  • loop of DNA. This is actually a plasmid but it looks the same way. You have a strand of

  • DNA in a perfect loop. And so for them they can just simply start copying it on this side.

  • The origin of replication is at one point. They move around and eventually what they'll

  • have is two strands of DNA. It's going to be an exact copy of that. And again in binary

  • fission those become different cells. But in us we have such a long DNA that we have

  • to wad it up to even get it to fit in a chromosome like these pictured right here. In other words

  • your DNA, in a cell is going to be like that long. And so if we were to start on one side

  • and start copying it, it would take forever. And so basically what happen is we work in

  • two directions. So basically there will be a site of replication where it starts here.

  • But we're going to have it moving in this direction and moving in that direction. And

  • so basically that diagram that I just showed you, I think this would be a better picture

  • of it. That diagram where we had the DNA here. And then we had those new strands of DNA that

  • are being formed. This would be one of those replication forks we call it. But there would

  • be another replication fork at the other side. And also in eukaryotic cells we'll have multiple

  • sites or multiple origins or replication. So we'll have one here. We'll have on here.

  • We'll have one here. In other words when we're copying the DNA it's going to start copying

  • in a bunch of different points. And then those replication forks will move towards each other

  • until we eventually have two strands of DNA. And so again, DNA replication is super important.

  • It's incredibly accurate. It rarely makes mistakes. And I hope that was helpful.

Hi. It's Mr. Andersen and in this podcast I'm going to talk about DNA replication.

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