00:05:24.06 We see that there's also 00:08:06.26 as early as the 1950s by Francis Crick, 00:06:28.20 we had termination factors, 00:10:07.21 What we found over time 00:39:31.14 We see that's true when we look at the way they bind to the ribosome. 00:00:21.24 to trigger a series of events in the cell 00:32:28.20 So, this is a global view of what 00:30:13.07 eIF5B, which is a homologue of IF2, 00:01:39.21 in the form of nucleotides, 00:06:25.10 are specified by just one codon. 00:31:05.20 So, at the heart of the elongation reaction 00:26:50.08 that are big, big targets that Dom34 needs to act on. 00:34:21.05 that joins two amino acids together, 00:24:51.04 And to us this looked like it might be a No-Go substrate. 00:02:29.26 grew the process of translation, 00:03:11.18 in these processes of decay, 00:33:33.22 And we see, for example, these adenosines here 00:37:56.18 and likely functions much like a pawl in a motor 00:17:00.04 The ribosome is the enzyme 00:13:56.26 And we reasoned that 00:34:53.29 of that intermediate state. 00:33:33.20 - the long reads and the short reads - 00:38:21.22 through an open reading frame, 00:20:44.16 And in fact that's exactly what we saw. 00:28:01.24 is then effectively positioned in the P site of the ribosome 00:08:33.04 in the context of how we understand translation now. 00:26:19.13 what do the initiation factors do 00:28:07.01 But what we wanted to know was: 00:32:20.26 and it's going to be more likely to go through the next step, 00:23:09.12 and we'll describe those. 00:14:47.01 and you can ask what the occupancy 00:14:56.08 where you randomly fragment all of the messenger RNAs in a cell 00:28:48.24 in fact do cause the ribosome to stutter a bit 00:17:30.26 after RNAse treatment, we see this black trace here... 00:32:44.04 And that's again based on the codon-anticodon pairing. 00:06:10.28 and we see there are three stops: 00:12:39.06 is extremely high: 00:03:01.04 getting rid of the messenger RNAs, 00:29:47.28 The next step in bacteria and eukaryotes 00:08:01.04 So, the idea of an adaptor 00:32:43.14 and far-reaching 00:33:40.28 on endonucleolytically-cleaved messenger RNAs 00:23:27.26 and the ideas we had going into this project. This field is for validation purposes and should be left unchanged. 00:25:19.15 when we look at this EM (electron microscopy) image of translation in a cell 00:20:53.12 more able to do complex reactions or complex regulation. 00:39:49.16 while they have similar names 00:36:00.25 the amino acid that's charged on it, 00:06:34.14 which is AUG. 00:16:59.23 So, we felt very confident 00:19:20.28 and therefore there's only 16 nucleotides there, 00:12:14.07 with other results in the field 00:18:59.08 in the process of translocating along a messenger RNA, 00:04:01.18 and there are some significant differences 00:29:52.14 We need the large subunit to come in. 00:18:27.11 We can see the distribution, here, 00:10:39.22 And what we've found is that 00:33:32.12 is being interpreted. 00:05:43.03 and we see that protein synthesis, 00:12:12.06 And at the heart of that process 00:21:12.14 but they likely are going to be 00:14:02.03 because occasionally the enzyme 00:24:26.16 and that ends up presenting some unique challenges 00:04:34.10 that decides whether a message is good or not. 00:11:03.02 for these proteins. 00:30:57.10 and it's a relatively fast process, 00:05:06.01 that's responsible for the catalysis of peptide release. Textbook 00:16:49.07 By contrast, the ribosome footprints 00:31:09.11 We see, trailing behind that signature... 00:10:52.18 when the messenger RNA template, here, tends to be short. 00:02:10.26 and I think that's the best way to begin thinking about 00:09:41.18 the T-loop and the D-loop. 00:04:17.24 that we had been studying, 00:15:07.20 because you know how much of each messenger RNA is present 00:25:08.01 happening in the cell is in the context of what we call a polysome, In translation, mRNA along with transfer RNA (tRNA) and ribosomes work together to produce proteins. 00:21:26.24 in the Dom34 deletion strain relative to a wild type strain. 00:21:15.22 and we might be able to decipher something 00:23:38.01 whereas these other eukaryotic-specific factors 00:26:27.20 really does about the same thing in both systems, 00:13:28.13 onto the end of the specific tRNAs that it recognizes. 00:03:40.06 where Roy's lab put into a messenger RNA in yeast 00:17:55.22 There's a number of proteins, 00:10:44.13 There tend to be 30-40 tRNAs in a cell. 00:28:04.11 and we're not quite sure what might happen to these ribosomes. 00:41:43.29 that evolved long before the divergence 00:02:59.12 nproductive messenger RNAs, 00:27:52.10 and it actually binds directly to it 00:36:20.26 that the incomplete protein product needs to be targeted for degradation, 00:28:34.12 - 95% of messenger RNAs - 00:36:51.00 within the context of the ribosome 00:08:22.17 to radioactively-labeled amino acids, 00:11:54.04 and this explains how 30-40 tRNAs 00:19:19.20 this is another view of the ribosome 00:11:31.24 and we refer to that as wobble. 00:00:11.18 and in the Howard Hughes Medical Institute, 00:34:09.01 of universally conserved nucleotides 00:08:41.24 the dipeptide Met-Phe. 00:38:44.10 and they have to have an end, actually, that promotes chemistry. Protein Synthesis Definition In Biology Top 15 Reasons to Avoid Low Carb Diets ...When insulin ranges are chronically as well minimal as they may wellbe in incredibly minimal carb diet programs, catabolism (breakdown) ofmuscle mass protein increases, and protein synthesis … 00:05:37.01 So, built into the code also has to be 00:30:28.04 Elongation is the process by which 00:03:14.06 iteratively adding an amino acid to the growing chain, 00:33:08.22 invisible by normal experiments. 00:20:56.15 that we understood something about ribosome functions 00:36:30.25 to perform a relatively simple chemical reaction, 00:06:52.19 and that's what we mean by non-ambiguous. 00:41:59.28 with the help of exogenous protein factors, of course. 00:33:03.09 that accelerate some of the forward rate constants in this scheme 00:11:11.08 Well, over the same time period 00:10:05.05 and this is sort of the summary of that story. 00:13:31.02 And so the question that we really wanted to ask next 00:32:00.02 those are the grey 30-nucleotide long reads. 00:29:12.02 They form a circular complex; 00:16:50.13 that are going to be evaluated by the protein synthesis machinery 00:21:05.13 we have an additional layer of RNA elements 00:15:24.08 and so this gives me an opportunity 00:25:29.00 and we see the ribosomes accumulating along it, 00:16:27.02 by key elements in the eukaryotic translation machinery. 00:29:56.22 and what suppressor tRNAs do 00:35:40.16 by working in a genetically-manipulable system 00:02:50.21 is focus on two general areas. 00:21:31.22 is you can take all the reads on a given gene 00:06:23.08 We also see that some amino acids 00:09:30.22 We call it the acceptor end. 00:00:35.22 The genetic information is found in the form of DNA, 00:18:17.22 It's just that the ribosome is going to facilitate that march. 00:12:06.12 leading to the recycling reaction 00:22:32.01 what I was focusing on was full-length reads 00:02:40.04 is that the ribosome senses that it's making lysine-lysine-lysine-lysine, 00:03:25.04 which is when, at the end of a coding region, 00:08:45.15 where we've got a tRNA that's been labeled, 00:37:15.09 really does take place. 00:17:10.04 I'm going to tell you that we added another little touch 00:02:53.00 and Non-Stop Decay 00:04:13.26 of the eukaryotic termination factors 00:19:33.14 to the decoding center 00:33:00.28 is largely erased by a variety of quality control pathways 00:22:55.09 just as IF1, IF2, and IF3, 00:19:58.08 is probably some natural pausing by ribosomes, 00:09:29.27 was actually resulting in the release 00:36:15.12 There's an element in the large subunit 00:36:07.16 and we think the main thing that the ribosome does 00:11:32.26 behind the ribosome, 00:26:49.10 and find its AUG 00:24:16.23 specifically when IRE1 endonuclease 00:04:03.20 that we'd like to discuss. 00:05:41.08 and how you end protein synthesis, 00:12:31.18 there are exactly 20 aminoacyl-tRNA synthetases in most cells. 00:39:36.03 on that side of the ribosome 00:08:08.15 who realized there had to be such a molecule 00:34:23.24 that the ribosome catalyzes. 00:15:38.17 of appropriate start sites in bacterial messenger RNAs, 00:25:27.01 spread out along this grid, 00:22:27.28 and it had to do with those short reads. 00:18:03.15 So, we isolated shorter and longer footprints as well, 00:02:00.12 If the messenger RNA picked up a cleavage 00:11:52.16 only at the third position, 00:26:54.16 which is that when ribosomes pause, 00:21:38.01 by which the ribosome is going to find the start codon Both protein synthesis and DNA replication can make errors by the incorporation of incorrect precursor molecules. 00:22:09.06 There is one exception, 00:34:28.29 So, I showed you in the beginning 00:27:24.10 in this Non-Stop-Decay pathway 00:11:22.15 there are perfect Watson-Crick pairing interactions 00:06:44.24 and other steps in the translation system. 00:35:10.16 there's plenty of short-mer reads in the cell 00:25:13.08 loaded onto a single messenger RNA, like this, Science 330: 369-72. 00:42:12.06 And in a number of points Scientists can also synthesize artificial peptides usi… 00:20:29.18 known as 3-AT - 3-aminotriazole - 00:09:15.26 to these two different complexes, 00:29:05.10 that poly-lysine is a problem. 00:08:47.29 We refer to it as a cloverleaf. 00:07:52.24 So, on the methionyl-tRNA, 00:18:36.07 New tRNAs are going to come into this site of the ribosome. 00:01:18.22 or a polymer of amino acids that we refer to as proteins. 00:39:25.17 the hydrolytic reaction, 00:19:36.29 And I think this is a nice view of the ribosome 00:00:13.04 and what I'm going to tell you about today 00:10:52.01 and we refer to an event known as wobble 00:04:41.26 can cover the 20 amino acid complexity, 00:34:06.10 as many as 50% of the genes that we look at, 00:21:30.01 The way that you can do this experiment 00:31:41.20 when you read into polyA tail. 00:08:28.19 it runs as a big 80S complex. 00:16:02.04 There's a periodicity to the footprint signature. 00:27:54.06 and effectively tethers the messenger RNA 00:41:19.14 the subunits are split, 00:34:55.29 this is actually a wild type strain 00:13:34.02 where we see the square amino acid, here, 00:09:49.22 and this activity suggested 00:31:46.16 what if we look at this in a more global way? 00:21:19.16 So, that's kind of an exciting way 00:12:37.07 Is is the same in wild type conditions 00:24:06.02 really function in both systems. 00:16:57.23 you can see that three nucleotide periodicity emerging. 00:30:55.04 where accuracy is imposed, 00:40:11.20 there really wasn't termination Identify the steps of transcription, and summarize what happens during each step. 00:29:54.06 n bacteria, that's facilitated 00:25:17.05 both for the disome reads 00:35:29.19 is formed exclusively, really, of RNA components, 00:12:15.20 that were taking place in intact cells. 00:09:46.10 a novel activity for Dom34 and Hbs1 proteins, 00:14:28.25 Well, let me tell you a little bit about the messenger RNAs. 00:02:48.25 So, what I'm going to do today 00:24:37.15 and follow how it is that ribosomes identify the initiating AUG 00:27:21.16 is this Shine-Dalgarno motif 00:11:28.16 is recognizing the UUU codon, 00:26:39.13 on this single gene, 00:34:57.18 that we're looking at here... 00:38:49.10 but hydrolysis. 00:07:46.18 and in the A site we've placed a stop codon, 00:13:58.15 that has the opportunity 00:10:22.18 So, an additional feature that needs to be discussed 00:34:26.10 how important we think Dom34 might be in the cell. 00:12:03.11 is how is it that this key step happens, 00:11:20.27 And it fit in with models he had presented, 00:29:18.23 What happens next is a ribosome complex, 00:38:50.16 They're going to bring a water into the active site 00:25:10.20 and the wild type ribosome footprints, 00:29:20.26 that's actually bound to those factors 00:24:02.19 that in yeast cells is exported from the nucleus 00:11:27.08 maybe, for example, a big stem-loop structure. 00:35:13.26 This is the interface side of the ribosome 00:08:24.12 and that those radioactively-labeled amino acids 00:18:13.11 or a Dom34 delta strain... 00:05:44.27 or we've learned that protein synthesis 00:26:40.20 We see that another initiation factor 00:31:54.06 to help evaluate the quality 00:10:27.26 was codon independent. 00:35:05.07 out of Tom Steitz's lab. 00:19:22.26 but these are principal targets 00:24:27.26 that looks like this, the HAC1 gene, It is the second part of the central dogma in genetics. 00:18:41.19 And tRNAs leaving the ribosome 00:02:57.26 at keeping under control in the cell u 00:35:38.26 and we've uncovered that role 00:34:50.16 and they might be typically targeted by the Dom34 system. 00:35:42.08 where we can actually delete the gene 00:08:44.00 On the left we see the typical 2-dimensional structure 00:14:43.20 You get hundreds of millions of reads In her first talk, Green provides a detailed look at protein synthesis, or translation. 00:40:09.22 So, that's a pretty exciting idea is that 00:01:34.14 they, by mutation of some of other process 00:04:32.05 and so any code that would comprehensively Protein synthesis is a biological procedure which living cells perform to create new proteins. 00:00:40.01 built of nucleotide building blocks, 00:12:26.15 behaved on reporters in various cells, 00:41:05.12 But the process in each case... 00:14:44.22 If we look on the left we see 00:07:34.08 For the one on the left, How exactly do all those letters in DNA code for making a living thing? 00:40:38.28 So finally, we're almost done. 00:23:06.18 ribosome occupancy on those short-mer fragments 00:20:16.26 in the absence of Dom34 that accumulated. 00:01:30.12 That is, instead of Examples of proteins include enzymes, antibodies and some hormones which help to speed up chemical reactions, defend against diseases and regulate the activity of cells.Proteins also play a role in movement, structural support, storage, communication between cells, digestion and the transport of substances around the body. 00:03:35.16 for a number of years. 00:04:09.05 Dom34 and Hbs1, 00:16:17.26 It's an unusual nucleotide 00:02:15.15 It would also like to get rid of that incomplete polypeptide product. 00:13:36.16 the orange amino acid, 00:23:02.17 So, these are essentially equivalent factors in bacteria and eukaryotes 00:36:44.06 So, now we've formed a peptide bond. 00:06:29.22 this UGG in the middle of the stop codon box, 00:13:43.08 is activated with ATP, 00:05:54.20 So, it turns out, in every open reading frame 00:09:06.06 that we refer to as the anticodon 00:33:54.26 knowing that this is a good interaction, 00:41:13.06 and an independent factor known as ABCE1, 00:23:36.26 and our increased ribosome occupancy 00:39:17.05 and it again suggests 00:01:32.10 having just a stop codon at the end of the open reading frame, 00:09:09.10 - not a deacylated tRNA band. 00:05:09.17 We see that Dom34 has similar domains. 00:20:52.04 throughout every gene in the genome. 00:00:31.20 - the proteins in the cell - 00:05:57.20 these homologues of the termination factors, 00:27:49.28 which is instead of running into a stem-loop here, 00:33:22.14 in fact using RNA sequencing methods, 00:36:13.10 It brings them together with universally conserved elements. 00:32:03.01 on all the genes 00:02:56.01 that are all aimed 00:06:06.19 We see that the code also specifies stops, 00:19:53.04 that's going to need to facilitate these events. 00:07:51.14 we see that the amino acid is labeled. 00:17:36.25 and that's where the exit channel exists, 00:26:05.16 is the formation of an initiation complex 00:42:23.25 which has a multi-step process 00:09:40.12 or free tRNA. The short answer is that the code of letters in DNA is a recipe to make proteins.Proteins form structures, enzymes or other chemicals to create the building blocks of organisms and maintain homeostasis. She then moved to Harvard to pursue her PhD in the lab of Jack Szostak where she worked on designing catalytic RNA molecules and investigating their implications for the evolution of life. Synthesis of the Protein from mRNA: Introduction, Steps, Essential parts, Significance. 00:12:13.23 is an enzyme known as the aminoacyl tRNA synthetase 00:42:04.17 are going to be the process of initiation 00:41:01.22 they're completely different factors 00:04:19.08 and we had been studying them biochemically. 00:03:02.04 about the process of translation, Your email address will not be published. 00:42:26.18 We know the fidelity of that process 00:13:23.29 by forming a chemical linkage, 00:30:53.17 and we'll talk about a number of steps 00:21:46.26 It's going to find the AUG 00:29:51.18 But the experiment we did While there is no definitive definition of a peptide, it usually refers to flexible (little secondary structure) chains of up to 30-50 amino acids. 00:18:06.02 so we were generous in slicing our fragments from a gel. 00:27:16.28 Alright, I'm going to tell you another story, now, 00:20:01.06 and cloning biases, Peptides may have their own biological function or be a structural and functional part of a protein molecule. 00:12:05.02 which is that the amino acids, By comparing protein synthesis in bacteria and eukaryotes, Green explains that it is possible to determine which components and steps are highly conserved and predate the divergence of different kingdoms on the tree of life, and which are more recent adaptations. 00:00:49.08 and with a polyA tail at the 3' end, 00:10:50.10 must recognize more than one codon, 00:18:47.18 We can see this in the context of 00:07:45.02 So, the next component that we're going to talk about is tRNA. 00:36:35.26 who did the work. 00:36:03.16 and that’s the attack of the nucleophile on the electron-deficient center. 00:29:39.08 and what we see is what we expect, It is then folded into the correct unique … 00:09:00.16 and it's on the anticodon stem, 00:41:24.04 and subunits for the next round of protein synthesis. 00:31:39.14 or into the decoding center. 00:33:26.11 in these two genes. Content. 00:01:04.18 which is the genetic information, 00:02:03.26 the ribosome would never reach a stop codon at the end, 00:23:20.25 you're actually enriched in those short-mer reads 00:15:30.16 from wild type yeast and from Dom34 mutant yeast 00:10:42.12 lacks the GGQ motif that's responsible for that activity 00:31:27.04 It protects the labile bond; 00:34:16.16 So, next I'm going to talk a little bit 00:28:08.23 was there any evidence of this sort of activity 00:02:50.21 So, Nonsense-Mediated Decay, 00:10:48.04 And a little aside is that these proteins 00:18:11.08 If we take, from a wild type strain 00:06:35.28 is to form ribosome complexes 00:07:38.17 has a relatively modest consequence. 00:34:46.11 and that funny position is going to be resolved 00:26:48.19 that are crisis, 00:05:02.01 that could cover, easily, the 20 amino acids, 00:01:07.24 that signifies the end of an open reading frame, 00:14:38.19 from all the ribosomes in the cell, 00:29:34.07 until it hits the AUG... Translation is the process by which nucleotides, the “language” of DNA and RNA, are translated into amino acids, the “language” of proteins. 00:18:18.08 and ask what the average length 00:24:54.04 It's an open reading frame 00:36:47.03 And I'd like to also thank my funding sources 00:25:26.28 so just one ribosome - 00:03:33.00 has to be recycled in order to begin again, 00:32:47.19 into this yeast strain, 00:04:05.14 Then we're going to talk about the catalyst of this event, 00:00:19.29 identifies bad messenger RNAs 00:30:54.14 to see signs of this endonucleolytic cleavage 00:03:00.20 And then we're going to talk sequentially 00:11:48.02 encounter that endonucleolytically-cleaved end, 00:27:36.24 actually has something that we call the anti-Shine-Dalgarno motif in it. 00:02:31.13 which is now a very sophisticated 00:33:38.05 within another helix, and that's where they are normally, 00:08:31.09 and this all makes sense 00:34:39.08 what you can see here was there was about 5% of the reads, 00:25:32.12 relative to the wild type strain, 00:40:13.08 before the split of these domains of life, 00:34:22.08 where I talked about the distribution of fragment sizes, 00:34:44.01 into a funny position after peptide bond formation 00:26:46.27 there are very few Dom34 targets 00:26:36.10 binds to the small subunit of the bacterial ribosome 00:22:24.01 that showed us something new and exciting 00:25:47.04 In bacteria, 00:36:09.23 that are responsible for recognizing and recruiting factors 00:19:35.02 where the genetic information is interpreted. 00:14:46.13 the elements of a bacterial messenger RNA, 00:02:37.02 PolyA encodes lysine, 00:37:48.10 and this is where the A site tRNA would be, 00:15:03.16 and then you can also ask 00:28:54.09 and that kind of propagates along the length 00:27:14.07 that seem to block the wrong sites on the ribosome, 00:26:52.27 But it provided evidence for the model, 00:20:40.06 ribosomes piled up at histidine codons 00:11:53.17 Dom34 and Hbs1. 00:32:49.04 and that's maybe not a general phenomenon. 00:37:02.08 It's a GTPase 00:07:22.01 which are two amino acids that are positively charged, 00:29:12.20 So, we had another way that we thought we could look for 00:31:12.15 - these are short-mer reads. 00:24:22.06 And finally, I'll say that, in the end, 00:13:12.24 that there are ribosome insufficiencies in the cell, 00:31:27.06 So, this ribosome encounters polyA tail, 00:02:49.08 that we refer to as non-stop decay. 00:19:47.14 and what we see is that in a Dom34 knockout (KO) strain 00:15:58.08 and that this methodology is able to capture 00:19:56.18 Included in this bounce 00:15:15.19 has an AUG associated with it, 00:22:47.10 is that there are some core initiation factors, She … 00:35:49.04 This is the peptidyl-tRNA, here. 00:20:50.14 exactly where there are histidine codons 00:24:12.28 but the factors that have evolved 00:01:18.24 in RNA processing or with mutations 00:08:55.12 of a tRNA. 00:00:58.14 and protein is composed of amino acids. 00:33:43.16 and a cognate tRNA, 00:28:27.08 or in the yeast transcriptome, 00:41:29.01 I hope you've learned a number 00:21:42.26 and it's going to happen, actually, 00:40:23.21 A particular feature to notice, however, 00:07:00.08 the tRNA substrates, 00:34:59.20 and that's emphasized by this view, here, 00:39:55.18 and they have a motif that does chemistry, 00:41:57.07 at the heart of all of these events 00:32:25.06 If it's a poor binder 00:35:54.03 This is the growing polypeptide chain. 00:10:37.20 but that leaves 61 codons. The actual process of protein synthesis takes place in the cell cytoplasm, and it occurs on multiple ribosomes simultaneously. 00:26:23.15 Well, I can tell you that the set of core factors 00:28:28.22 and we can actually do sort of averaged gene 00:35:22.07 why this gene is essential in higher eukaryotes, 00:15:48.15 that the messenger RNA we're showing here 00:38:28.00 the genetic code specifies three different stop codons. 00:02:38.19 sort of in its simplicity, 00:29:01.12 This scanning takes place 00:28:35.01 or genes that have two lysines in a row 00:00:09.14 and I'm at the Johns Hopkins University School of Medicine 00:28:51.10 - you've got bigger peaks here, 00:31:01.10 in the same strain with the nonsense-suppressor tRNA 00:30:18.27 and the many, many initiation factors leave. 00:17:38.16 where the polypeptide emerges 00:09:03.19 and we saw the appearance of a different band, 00:25:51.04 and that's really the key to finding the AUG in bacteria. 00:32:07.18 What we see is the pile of grey reads right here 00:14:43.00 when they're ready for protein synthesis. My name is Rachel Green. 00:36:27.16 to bring these two substrates together A living cell has the ability to synthesize hundreds of separate proteins per second. 00:10:56.28 to explain how one tRNA 00:26:09.26 with an initiator tRNA bound in the P site of the ribosome, 00:32:17.28 And if it's a nice tight interaction, 00:25:33.16 is being translated. 00:26:01.02 we saw another signature of Dom34 activity in the cell 00:14:23.19 you can digest away all the free messenger RNA, 00:23:12.18 consistent with the idea that Dom34 00:30:15.02 these are full-length reads we're looking at now... 00:05:40.08 and we suspected must engage the ribosome, 00:29:45.16 It's found the AUG and it's ready to go. 00:12:00.06 And what these factors might allow 00:10:36.02 that are not recognized by tRNAs, 00:04:26.10 is in fact ribosome-driven, as you might suspect, 00:07:09.06 in a way that's conservative, 00:41:58.16 there's RNA driving these steps, 00:23:12.28 of eukaryotic-specific factors 00:19:08.08 are these truncated fragments down here 00:23:33.08 what I want to show you next 00:07:34.24 These are two negatively charged amino acids, 00:30:30.21 in the Dom34 mutant. 00:00:55.08 that tells a ribosome 00:22:06.04 by the machinery specifically responsible 00:35:03.13 This is a crystal structure that came originally 00:01:00.12 Also, we know that the typical messenger RNAs 00:37:10.19 might be to insert something 00:23:00.02 eIF1A, eIF5B, and eIF1. 00:10:41.20 there are never as many as 61 tRNAs in any given cell. 00:28:24.02 and we can do that by looking at all of the ribosomes in the yeast genome, 00:18:36.07 what we see is that a majority of our reads 00:31:57.13 of the codon-anticodon interaction 00:32:37.14 in this Dom34-delta strain. 00:12:58.08 We did, however, know that the Dom34 deletion in yeast 00:10:19.16 that's going to read the codon in the messenger RNA template. 00:20:07.19 is in this particular gene 00:04:57.21 called the NIKS domain, 00:31:41.10 That process is outlined here, 00:20:48.10 and you can see that there's addition of protein and RNA components 00:13:26.27 that that's when cells really get sick, 00:28:53.11 starts at the 5' end of a messenger RNA 00:41:51.27 at the heart of peptidyl transfer, 00:15:21.09 which is the ending site, 00:33:46.00 of the ribosome reading into poly-lysine. 00:35:12.16 and you could imagine there's short-mers 00:07:42.26 or the genetic code. The order in which amino acids are joined together determine the shape, properties, and function of a protein. 00:15:22.23 and they have this interesting element upstream, 00:31:38.12 Again, identifying a clear role for Dom34 00:25:17.05 and we can see how this might look in a cell 00:19:05.20 Finally, the fragments that we were most interested in 00:33:36.21 we see a clear signature 00:22:16.24 to allow for the next round of translation. 00:17:55.17 might actually be also biased 00:07:20.03 If we compare lysine and arginine, 00:07:19.26 about Dom34 and Hbs1 function, 00:35:59.16 is really that at the center 00:14:40.10 you can submit it for high-throughout sequencing. 00:25:01.17 We can look at our ribosome profiling data 00:25:11.15 which is multiple ribosomes 00:02:06.12 and the ribosome would be stuck 00:09:02.12 and what this portion of the molecule does 00:35:02.11 of the ribosome. 00:22:42.21 which is, if we look at a wild type 00:03:39.26 So, the players... what are we gonna focus on first? 00:14:22.07 So, now we have activated aminoacyl-tRNAs. 00:07:56.17 We formed a similar complex on the other side, here, 00:02:57.08 the proteins, the ribosomes... 00:03:53.03 that encodes the genetic information. 00:00:44.29 has to be transcribed into a different form 00:13:54.19 on their various mRNA templates. 00:02:27.27 presumably in a primordial world, 00:05:29.20 if we look here at the top left corner, We found was that this reaction by Dom34 Okay, now for the aminoacyl.!, for the final steps: replication, transcription, the right amino acid 00:12:48.09 gets to. 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