1 00:00:14,760 --> 00:00:09,959 for Speaker of this session is me I am 2 00:00:16,859 --> 00:00:14,770 me I am from Georgia Tech I work kind of 3 00:00:18,870 --> 00:00:16,869 co advised between Nick head and Lauren 4 00:00:22,019 --> 00:00:18,880 Williams we're really lucky Georgia Tech 5 00:00:25,019 --> 00:00:22,029 to have a lot of funding for origins of 6 00:00:26,880 --> 00:00:25,029 life research so quick plug so I'm 7 00:00:29,040 --> 00:00:26,890 working in the center for ribosomal 8 00:00:30,870 --> 00:00:29,050 orange enough evolution so that's why 9 00:00:32,970 --> 00:00:30,880 today I'm going to be talking about what 10 00:00:34,800 --> 00:00:32,980 I'm what's probably going to drive some 11 00:00:38,939 --> 00:00:34,810 of the rock people in here crazy which 12 00:00:41,430 --> 00:00:38,949 is molecular paleontology so in this 13 00:00:42,810 --> 00:00:41,440 case we're kind of working on a top-down 14 00:00:46,080 --> 00:00:42,820 approach where we're trying to use 15 00:00:48,119 --> 00:00:46,090 what's left in modern biology and drill 16 00:00:49,560 --> 00:00:48,129 back as far as we can into evolutionary 17 00:00:52,470 --> 00:00:49,570 history with that and it turns out 18 00:00:55,319 --> 00:00:52,480 luckily we can drill back pretty far so 19 00:00:56,849 --> 00:00:55,329 you guys are already generally familiar 20 00:00:59,880 --> 00:00:56,859 with the ribosome you about to become 21 00:01:01,319 --> 00:00:59,890 intimately familiar with the ribosome so 22 00:01:05,009 --> 00:01:01,329 you guys have already seen the central 23 00:01:07,050 --> 00:01:05,019 dogma this is where the ribosome fits in 24 00:01:10,080 --> 00:01:07,060 here so the ribosome is what turns RNA 25 00:01:13,310 --> 00:01:10,090 into protein it is in terms of 26 00:01:17,969 --> 00:01:13,320 biomolecules huge it's one mega dalton 27 00:01:19,920 --> 00:01:17,979 1000 1,000,000 Dalton's it is as I 28 00:01:22,080 --> 00:01:19,930 already mentioned earlier probably well 29 00:01:23,999 --> 00:01:22,090 established maybe not in its full modern 30 00:01:25,139 --> 00:01:24,009 form but in a pretty advanced forum at 31 00:01:29,370 --> 00:01:25,149 the time of the last Universal common 32 00:01:31,889 --> 00:01:29,380 ancestor this is generally how it works 33 00:01:35,010 --> 00:01:31,899 in a sort of schematic way where it 34 00:01:38,100 --> 00:01:35,020 reads the messenger RNA and these tRNA 35 00:01:41,609 --> 00:01:38,110 serve as adapter molecules that turn 36 00:01:46,380 --> 00:01:41,619 that code into the specific amino acid 37 00:01:50,700 --> 00:01:46,390 that that code codes for and then you 38 00:01:53,279 --> 00:01:50,710 make a protein note of that this is what 39 00:01:55,230 --> 00:01:53,289 it looks like in more depth so here we 40 00:01:58,080 --> 00:01:55,240 have separated out the small subunit 41 00:02:02,179 --> 00:01:58,090 which is responsible for binding the 42 00:02:05,310 --> 00:02:02,189 messenger RNA and it is comprised of 43 00:02:08,460 --> 00:02:05,320 what's called the 16s ribosomal RNA and 44 00:02:09,840 --> 00:02:08,470 about 20 ribosomal proteins but I don't 45 00:02:11,100 --> 00:02:09,850 care about that this morning I'm going 46 00:02:13,949 --> 00:02:11,110 to talk to you solely about the large 47 00:02:15,990 --> 00:02:13,959 subunit which is responsible for the 48 00:02:17,670 --> 00:02:16,000 peptide bond formation so it's pretty 49 00:02:19,190 --> 00:02:17,680 widely accepted that the small subunit 50 00:02:21,380 --> 00:02:19,200 came later 51 00:02:22,850 --> 00:02:21,390 Lucien so we're trying to go back as far 52 00:02:25,460 --> 00:02:22,860 as we can so we're just going to look at 53 00:02:29,479 --> 00:02:25,470 the large subunit so it's where the 54 00:02:33,800 --> 00:02:29,489 actual chemistry happens so the 23s and 55 00:02:36,140 --> 00:02:33,810 5s ribosomal RNAs are the RNA components 56 00:02:37,699 --> 00:02:36,150 of the large subunit again I'm not going 57 00:02:38,960 --> 00:02:37,709 to talk about the 5s because it's much 58 00:02:42,410 --> 00:02:38,970 later we're going to talk solely about 59 00:02:43,729 --> 00:02:42,420 the 23s RNA and a subset of these 30 60 00:02:47,839 --> 00:02:43,739 proteins so when you're looking at this 61 00:02:49,220 --> 00:02:47,849 this here's the 5s RNA I will generally 62 00:02:51,020 --> 00:02:49,230 tell you today that if something looks 63 00:02:53,839 --> 00:02:51,030 like it's on the exterior it's probably 64 00:02:55,220 --> 00:02:53,849 newer so the 5s RNA can see is right out 65 00:02:58,009 --> 00:02:55,230 of here on the outside probably a lot 66 00:03:00,289 --> 00:02:58,019 newer addition to the ribosome and then 67 00:03:02,839 --> 00:03:00,299 these beige things here that look like 68 00:03:05,210 --> 00:03:02,849 hela sees those are the right the 23s 69 00:03:06,830 --> 00:03:05,220 and the 16s ribosomal RNAs these purple 70 00:03:08,120 --> 00:03:06,840 guys that are sort of stuck in all over 71 00:03:10,550 --> 00:03:08,130 the surface those are the ribosomal 72 00:03:12,550 --> 00:03:10,560 proteins so this in particular is a 73 00:03:18,319 --> 00:03:12,560 bacterial ribosome you care what 74 00:03:21,770 --> 00:03:18,329 ribosomes are even bigger so what we 75 00:03:24,650 --> 00:03:21,780 wanted to do was try and resurrect an 76 00:03:26,270 --> 00:03:24,660 ancestral version of the ribosome so 77 00:03:28,640 --> 00:03:26,280 what we call it was the ancestral 78 00:03:31,550 --> 00:03:28,650 peptidyl transferase center a ptc for 79 00:03:33,050 --> 00:03:31,560 short and basically because that's what 80 00:03:34,640 --> 00:03:33,060 we wanted to do we want it to be able to 81 00:03:37,520 --> 00:03:34,650 make peptide bonds to do the peptidyl 82 00:03:39,770 --> 00:03:37,530 transferase reaction but in a much more 83 00:03:42,470 --> 00:03:39,780 simplified way than we have today so 84 00:03:45,289 --> 00:03:42,480 included in our model are the ancestral 85 00:03:47,720 --> 00:03:45,299 ribosomal RNA it's a very stripped down 86 00:03:49,970 --> 00:03:47,730 version of the 23s RNA which i'll show 87 00:03:52,610 --> 00:03:49,980 you on the next slide and it's stitched 88 00:03:54,140 --> 00:03:52,620 together with these tetra loops which 89 00:03:55,340 --> 00:03:54,150 all you need to know about those is that 90 00:03:57,890 --> 00:03:55,350 they're very stable we know that they 91 00:03:59,569 --> 00:03:57,900 fold in a very predictable way so 92 00:04:02,890 --> 00:03:59,579 they're kind of anywhere that we made a 93 00:04:04,879 --> 00:04:02,900 cut in the RNA we put those in and five 94 00:04:06,920 --> 00:04:04,889 ribosomal peptides that are highly 95 00:04:09,110 --> 00:04:06,930 conserved and penetrate very deeply down 96 00:04:12,170 --> 00:04:09,120 into the core of the ribosome those are 97 00:04:14,629 --> 00:04:12,180 derived from ribosomal proteins large 98 00:04:17,300 --> 00:04:14,639 subunit ribosome proteins l2 l3 l4 l5 99 00:04:18,920 --> 00:04:17,310 teen and l 22 all you really need to 100 00:04:20,449 --> 00:04:18,930 know about those is that there's a bunch 101 00:04:24,500 --> 00:04:20,459 of them this is just a few of them and 102 00:04:26,180 --> 00:04:24,510 that the L stands for large subunit so 103 00:04:29,330 --> 00:04:26,190 in this case in the background here you 104 00:04:30,830 --> 00:04:29,340 can see the entire 23s ribosomal RNA for 105 00:04:32,450 --> 00:04:30,840 comparison so you can see how much we 106 00:04:34,189 --> 00:04:32,460 really have stripped away 107 00:04:39,170 --> 00:04:34,199 in this really it's about eighty percent 108 00:04:40,670 --> 00:04:39,180 of the RNA and so these are the co 109 00:04:42,020 --> 00:04:40,680 crystallized logins from this crystal 110 00:04:43,640 --> 00:04:42,030 structure that just give you an idea of 111 00:04:46,339 --> 00:04:43,650 where in this beast the actual chemistry 112 00:04:50,480 --> 00:04:46,349 happens so it's right down in here right 113 00:04:53,390 --> 00:04:50,490 in the center so this is what the 114 00:04:54,920 --> 00:04:53,400 secondary structure of the 23s ribosomal 115 00:04:58,370 --> 00:04:54,930 RNA looks like secondary structure 116 00:05:00,379 --> 00:04:58,380 rather than this here which is a 3d 117 00:05:03,230 --> 00:05:00,389 structure is basically just laid flat 118 00:05:06,529 --> 00:05:03,240 for simplicity's sake so anywhere that 119 00:05:07,730 --> 00:05:06,539 there's a sort of two lines close to 120 00:05:09,439 --> 00:05:07,740 each other that means that there's base 121 00:05:12,529 --> 00:05:09,449 pairing going in between going between 122 00:05:14,990 --> 00:05:12,539 those two strands of the RNA so in the 123 00:05:18,200 --> 00:05:15,000 dash line here in the back that's the 124 00:05:21,110 --> 00:05:18,210 entire 23s ribosomal RNA in the black is 125 00:05:22,610 --> 00:05:21,120 in the sort of glue is what we kept so 126 00:05:24,499 --> 00:05:22,620 you can see again we'd stripped a lot of 127 00:05:27,110 --> 00:05:24,509 this away we made actually made this 128 00:05:29,330 --> 00:05:27,120 part in the lab which is it represents 129 00:05:33,770 --> 00:05:29,340 about twenty percent of a bacterial 130 00:05:35,629 --> 00:05:33,780 large subunit RNA and made it with some 131 00:05:37,270 --> 00:05:35,639 nice pcr techniques that I wish I had 132 00:05:40,790 --> 00:05:37,280 the time tell you about but I don't and 133 00:05:42,560 --> 00:05:40,800 these ribosomes peptides which I can 134 00:05:44,420 --> 00:05:42,570 tell you about more in a couple slides 135 00:05:47,570 --> 00:05:44,430 but first we need to tell you about my 136 00:05:50,510 --> 00:05:47,580 methods so a main method that I use is 137 00:05:51,589 --> 00:05:50,520 called shape and that stands actually 138 00:05:53,540 --> 00:05:51,599 Jessica showed some of this yesterday 139 00:05:55,520 --> 00:05:53,550 but it stands for selectively two prime 140 00:05:57,589 --> 00:05:55,530 hydroxyl isolation analyzed by primer 141 00:06:01,520 --> 00:05:57,599 extension astrobiologist and their 142 00:06:03,529 --> 00:06:01,530 acronyms so the way that it works in a 143 00:06:06,050 --> 00:06:03,539 very basic way is you have a chemical 144 00:06:09,140 --> 00:06:06,060 that selectively modifies your RNA at 145 00:06:11,120 --> 00:06:09,150 positions where it is flexible ie not 146 00:06:14,540 --> 00:06:11,130 base paired so what you get in the end 147 00:06:18,110 --> 00:06:14,550 is a readout of the places in your RNA 148 00:06:22,040 --> 00:06:18,120 that are that are not based paired and 149 00:06:26,180 --> 00:06:22,050 then from there we do reverse 150 00:06:29,060 --> 00:06:26,190 transcription into you just using an 151 00:06:31,730 --> 00:06:29,070 enzyme we hijack biology basically to 152 00:06:33,290 --> 00:06:31,740 make the complementary DNA that stops 153 00:06:35,810 --> 00:06:33,300 when it hits one of these modifications 154 00:06:38,839 --> 00:06:35,820 so what we're left with is a library of 155 00:06:41,089 --> 00:06:38,849 pieces of DNA that represent the places 156 00:06:43,579 --> 00:06:41,099 that are flexible in the corresponding 157 00:06:45,500 --> 00:06:43,589 RNA we run that on capillary 158 00:06:47,150 --> 00:06:45,510 electrophoresis which if you know any 159 00:06:50,690 --> 00:06:47,160 with gel electrophoresis is we act the 160 00:06:53,420 --> 00:06:50,700 same but its run in a capillary and from 161 00:06:55,370 --> 00:06:53,430 there we can process that data should we 162 00:06:57,410 --> 00:06:55,380 use in-house MATLAB script so we've 163 00:06:59,300 --> 00:06:57,420 developed to get a single nucleotide 164 00:07:04,700 --> 00:06:59,310 readout of the flexibility of that piece 165 00:07:07,460 --> 00:07:04,710 of RNA so we did that on this ancestral 166 00:07:09,920 --> 00:07:07,470 piece of ribosomal RNA because we wanted 167 00:07:11,180 --> 00:07:09,930 to see basically if regardless of the 168 00:07:13,250 --> 00:07:11,190 fact that we stripped away about eighty 169 00:07:17,120 --> 00:07:13,260 percent of the RNA if it still folds in 170 00:07:19,970 --> 00:07:17,130 a similar way and it does what you can 171 00:07:21,710 --> 00:07:19,980 see here is that anywhere that there's 172 00:07:24,320 --> 00:07:21,720 these triangles that means that it is 173 00:07:25,520 --> 00:07:24,330 flexible and those triangles line up 174 00:07:27,290 --> 00:07:25,530 really well with the single-stranded 175 00:07:30,920 --> 00:07:27,300 regions which we expect to be flexible 176 00:07:33,500 --> 00:07:30,930 so we were very happy about that and in 177 00:07:35,720 --> 00:07:33,510 this case I want to highlight that this 178 00:07:38,660 --> 00:07:35,730 was done just with the RNA on its own 179 00:07:40,340 --> 00:07:38,670 and some monovalent cations which just 180 00:07:42,890 --> 00:07:40,350 help to neutralize the backbone so that 181 00:07:44,900 --> 00:07:42,900 you get secondary structure formation 182 00:07:48,110 --> 00:07:44,910 you get the Gila sees actually able to 183 00:07:50,660 --> 00:07:48,120 form so no divalent cations or no none 184 00:07:52,670 --> 00:07:50,670 of the ribosome peptides we did have 185 00:07:54,650 --> 00:07:52,680 some hints early on that certain parts 186 00:07:56,540 --> 00:07:54,660 of the secondary structure that we were 187 00:07:58,820 --> 00:07:56,550 working with were not great this is 188 00:08:01,160 --> 00:07:58,830 going to be a little bit of an aside but 189 00:08:03,500 --> 00:08:01,170 for instance this region here I would 190 00:08:05,030 --> 00:08:03,510 have expected these nucleotides to be 191 00:08:06,950 --> 00:08:05,040 reactive but they were not but when I 192 00:08:08,990 --> 00:08:06,960 went back and looked at the actual 3d 193 00:08:13,070 --> 00:08:09,000 crystal structure it looked a little bit 194 00:08:16,010 --> 00:08:13,080 more like this where my data lines up a 195 00:08:20,930 --> 00:08:16,020 lot better and I was happy again so we 196 00:08:22,580 --> 00:08:20,940 had hints early on the 23s secondary 197 00:08:25,370 --> 00:08:22,590 structure which we derived this 198 00:08:27,920 --> 00:08:25,380 representation from was not perfect and 199 00:08:30,620 --> 00:08:27,930 so some guys in our lab decided to take 200 00:08:34,219 --> 00:08:30,630 that and run with it and rewrite the 201 00:08:35,390 --> 00:08:34,229 ribosome bible essentially so that big 202 00:08:39,260 --> 00:08:35,400 secondary structure i showed you the 203 00:08:41,089 --> 00:08:39,270 beginning is largely the way that it 204 00:08:44,180 --> 00:08:41,099 looks because of historical purposes and 205 00:08:47,000 --> 00:08:44,190 as young scientists we all know how much 206 00:08:49,670 --> 00:08:47,010 we hate that sort of stuff so in this 207 00:08:51,950 --> 00:08:49,680 case they basically redrew the map so 208 00:08:53,690 --> 00:08:51,960 that it looked so that it didn't have a 209 00:08:55,400 --> 00:08:53,700 gap between the two halves for starters 210 00:08:57,290 --> 00:08:55,410 and so that it represented more 211 00:08:58,880 --> 00:08:57,300 accurately the base pairing that's seen 212 00:08:59,480 --> 00:08:58,890 in the crystal structure so in 213 00:09:02,360 --> 00:08:59,490 particular 214 00:09:04,519 --> 00:09:02,370 it's all just brought together and this 215 00:09:06,019 --> 00:09:04,529 domain here which we call domain 0 216 00:09:10,340 --> 00:09:06,029 because they'd already named the others 217 00:09:14,510 --> 00:09:10,350 1 through 6 little facetious maybe but 218 00:09:16,250 --> 00:09:14,520 so is really makes up the core of the 219 00:09:20,210 --> 00:09:16,260 ribosome so what you're seeing here in 220 00:09:24,010 --> 00:09:20,220 blue is the ancestral ribosomal RNA just 221 00:09:30,260 --> 00:09:27,650 so going back to the ribosomal protein / 222 00:09:32,420 --> 00:09:30,270 peptides this is what the ribosomal 223 00:09:34,100 --> 00:09:32,430 proteins in general look like they have 224 00:09:37,220 --> 00:09:34,110 these big globular domains and these 225 00:09:38,780 --> 00:09:37,230 long fingers that have sort of really no 226 00:09:43,190 --> 00:09:38,790 structure that penetrate down deeply 227 00:09:45,470 --> 00:09:43,200 into the ribosome they vary in size 228 00:09:48,190 --> 00:09:45,480 depending on how far they penetrate and 229 00:09:52,400 --> 00:09:48,200 how much of it actually penetrates from 230 00:09:54,170 --> 00:09:52,410 18 amino acids to 45 this is what they 231 00:09:57,550 --> 00:09:54,180 look like if we look at the crystal 232 00:10:01,280 --> 00:09:57,560 structure of the in social ribosomal RNA 233 00:10:04,069 --> 00:10:01,290 with these individual peptides so you 234 00:10:05,480 --> 00:10:04,079 can see that they're kind of peppered 235 00:10:07,010 --> 00:10:05,490 around in some cases they get pretty 236 00:10:08,930 --> 00:10:07,020 close to the very middle which is where 237 00:10:10,480 --> 00:10:08,940 the chemistry happens but for the most 238 00:10:13,040 --> 00:10:10,490 part what these are here for is to 239 00:10:15,160 --> 00:10:13,050 stabilize the background the backbone so 240 00:10:17,540 --> 00:10:15,170 that you can get these negative charged 241 00:10:20,060 --> 00:10:17,550 are pieces of RNA coming closely 242 00:10:22,630 --> 00:10:20,070 together so previously we've been able 243 00:10:26,199 --> 00:10:22,640 to show that four of those five 244 00:10:29,660 --> 00:10:26,209 ribosomal peptides do actually bind our 245 00:10:31,340 --> 00:10:29,670 ribosomal RNA I don't have time to go 246 00:10:33,470 --> 00:10:31,350 into this but if you'd like to discuss 247 00:10:35,180 --> 00:10:33,480 it later I'd be happy to is this is a 248 00:10:41,090 --> 00:10:35,190 bunch of experiments that I didn't do 249 00:10:43,910 --> 00:10:41,100 but the only one that didn't was l2 but 250 00:10:45,949 --> 00:10:43,920 we didn't know if it was binding in the 251 00:10:47,720 --> 00:10:45,959 exact place that we wanted it to so 252 00:10:49,550 --> 00:10:47,730 that's where my research came in and I 253 00:10:51,230 --> 00:10:49,560 went through and used the crystal 254 00:10:53,569 --> 00:10:51,240 structure to say hey where's where these 255 00:10:55,970 --> 00:10:53,579 things actually are where do we want 256 00:10:57,260 --> 00:10:55,980 them to bind and in the secondary 257 00:10:59,690 --> 00:10:57,270 structure it looks something like this 258 00:11:01,400 --> 00:10:59,700 just in the different colors are the 259 00:11:03,590 --> 00:11:01,410 different locations that we might expect 260 00:11:07,760 --> 00:11:03,600 to see contacts if they're binding at 261 00:11:11,090 --> 00:11:07,770 the expected places this is what my raw 262 00:11:13,369 --> 00:11:11,100 shape data looks at looks like I'm not 263 00:11:16,219 --> 00:11:13,379 going to try and help you guys in 264 00:11:17,929 --> 00:11:16,229 too much except to know that anywhere so 265 00:11:19,279 --> 00:11:17,939 that any location with an error here 266 00:11:22,639 --> 00:11:19,289 that you kind of see a peak that 267 00:11:24,979 --> 00:11:22,649 increases or decreases so there's some 268 00:11:26,989 --> 00:11:24,989 that that appear as we increase the 269 00:11:30,139 --> 00:11:26,999 amount of these peptides in this case 270 00:11:32,029 --> 00:11:30,149 it's the l4 peptide that means that 271 00:11:35,689 --> 00:11:32,039 there is a change in the structure at 272 00:11:37,249 --> 00:11:35,699 that location you know it's a local in a 273 00:11:39,259 --> 00:11:37,259 lot of cases just a single nucleotide 274 00:11:41,179 --> 00:11:39,269 that's maybe becoming less flexible so 275 00:11:45,109 --> 00:11:41,189 maybe more lockdown or becoming more 276 00:11:47,869 --> 00:11:45,119 flexible and floppy when so far I've 277 00:11:50,299 --> 00:11:47,879 done l4 and I'll 22 peptides and when I 278 00:11:51,769 --> 00:11:50,309 compare where those actual changes 279 00:11:53,539 --> 00:11:51,779 happen this is in a very rough way I 280 00:11:56,109 --> 00:11:53,549 haven't fully processes data yet because 281 00:11:58,460 --> 00:11:56,119 it's kind of a nightmare to process but 282 00:12:00,559 --> 00:11:58,470 if I sort of do a really rough 283 00:12:04,639 --> 00:12:00,569 estimation these are the positions that 284 00:12:08,149 --> 00:12:04,649 I see some of the biggest changes at and 285 00:12:10,219 --> 00:12:08,159 so for the l4 in particular let's say 286 00:12:12,589 --> 00:12:10,229 this is one of the places that i do see 287 00:12:14,960 --> 00:12:12,599 a change and that is one of the places 288 00:12:16,579 --> 00:12:14,970 that i expect to sorry down here that i 289 00:12:18,919 --> 00:12:16,589 expect to see a change so very close 290 00:12:22,309 --> 00:12:18,929 which makes me very happy and again 291 00:12:25,129 --> 00:12:22,319 right here there's some not too far away 292 00:12:27,049 --> 00:12:25,139 that i expected to have a change in in 293 00:12:30,019 --> 00:12:27,059 their structures so what i can say right 294 00:12:33,379 --> 00:12:30,029 now is that both the l4 and l5 tides do 295 00:12:36,199 --> 00:12:33,389 induce modest changes in the second in 296 00:12:38,090 --> 00:12:36,209 the structure and in some cases at 297 00:12:41,449 --> 00:12:38,100 locations near where we expected them to 298 00:12:44,479 --> 00:12:41,459 and l3 alone I didn't show because it's 299 00:12:46,189 --> 00:12:44,489 boring it doesn't seem to bind at least 300 00:12:48,319 --> 00:12:46,199 in a specific way so it may still bind 301 00:12:50,419 --> 00:12:48,329 but just not the exact way that we 302 00:12:51,679 --> 00:12:50,429 intended it may be that it's dependent 303 00:12:54,229 --> 00:12:51,689 on having one of those other peptides 304 00:12:56,119 --> 00:12:54,239 there first so that you know you need to 305 00:12:58,909 --> 00:12:56,129 have one so that the other one can kind 306 00:13:01,129 --> 00:12:58,919 of cooperatively come in and in bind and 307 00:13:03,019 --> 00:13:01,139 in the future we're going to start doing 308 00:13:05,989 --> 00:13:03,029 these in combinations so that we can get 309 00:13:07,579 --> 00:13:05,999 an idea if there's an actual order of 310 00:13:08,989 --> 00:13:07,589 assembly and that sort of thing and also 311 00:13:11,779 --> 00:13:08,999 start adding back in stuff like 312 00:13:15,579 --> 00:13:11,789 magnesium which is also very important 313 00:13:18,139 --> 00:13:15,589 in forming RNA large RNA structures and 314 00:13:19,689 --> 00:13:18,149 that's everything I had these are all 315 00:13:22,849 --> 00:13:19,699 the people that helped with my work and 316 00:13:25,129 --> 00:13:22,859 once again thank NASA for funding us and 317 00:13:28,879 --> 00:13:25,139 for finding this conference in a large 318 00:13:30,349 --> 00:13:28,889 part and also the Center for ribosomal 319 00:13:43,639 --> 00:13:30,359 origins and evolution I will happily 320 00:13:50,349 --> 00:13:43,649 take any questions Bradley I see you 321 00:13:53,960 --> 00:13:50,359 have a question it's better be good oh 322 00:13:56,840 --> 00:13:53,970 it's awesome all right so you mentioned 323 00:13:59,389 --> 00:13:56,850 your future studies doing magnesium 324 00:14:02,269 --> 00:13:59,399 adding it there yeah you see where I'm 325 00:14:03,769 --> 00:14:02,279 going any temptation to try it with iron 326 00:14:07,069 --> 00:14:03,779 since I know your labs have the 327 00:14:09,319 --> 00:14:07,079 capability yes yeah that's definitely a 328 00:14:13,299 --> 00:14:09,329 down the road sort of thing too yeah 329 00:14:16,429 --> 00:14:13,309 because because iron has been shown to 330 00:14:18,710 --> 00:14:16,439 really mimic magnesium in terms of the 331 00:14:21,019 --> 00:14:18,720 way that it interacts with RNA so yes 332 00:14:22,699 --> 00:14:21,029 for certain I would like to do that part 333 00:14:25,099 --> 00:14:22,709 of it is just instrumentation issues we 334 00:14:26,689 --> 00:14:25,109 are in the process of getting an 335 00:14:30,019 --> 00:14:26,699 anaerobic chamber that will allow us to 336 00:14:32,359 --> 00:14:30,029 do it but we've got it from somebody 337 00:14:35,479 --> 00:14:32,369 else who cut it in half and tried to put 338 00:14:36,649 --> 00:14:35,489 it back together and so it leaks so 339 00:14:38,149 --> 00:14:36,659 we're getting the manufacturers to fix 340 00:14:39,649 --> 00:14:38,159 it but once we have that we should be 341 00:14:41,629 --> 00:14:39,659 able to do all kinds more interesting