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[Music]

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[Applause]

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good afternoon everyone

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so over past hour we've been through

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this journey with insertions

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duplications by luke insertions by

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george fox and the common core by

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Jessica and those patterns have been

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quite persistent over for billions of

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years and we've thought everything is

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sort of holes in the same way and in the

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next 15 or so minutes I want to invite

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you to a journey into a world when all

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this is no longer true and this

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Wonderland is named mitochondria so I'm

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going to be talking about structures of

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mitochondrial ribosomes and this work

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was done in collaboration with Alexia

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moons and Alan brown from skylight flap

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and Harvard Medical School in Stockholm

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University and all this research was

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supported by a Swedish Research Council

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and NASA Center for the origins of life

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so I think for funding and so as I said

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so this emergence of mitochondria was

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dated about two billions of years ago

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when an alpha proteobacteria basically

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gotten incorporated in version 8 it into

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a proto eukaryotic cell and now it's

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been a hypothesis proposed by lynn

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margulis about 50 years ago and this is

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now almost unanimously accepted by the

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scientific community the fact that the

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mitochondria has evolved from an alpha

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proteobacteria so and what happened upon

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this process there are many many many

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complicated different phenomena related

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to mutual gene transfer from

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mitochondria into a nucleus as well as

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protein experts

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from cytosol into mitochondria so as a

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result of all these processes

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mitochondria eventually lost its

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independence and it just now became an

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organelle and nevertheless the genomic

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analyses performed in 90s by Michael

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Gray had indicated that there are a

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number of genomes that are always

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retained within mitochondria among them

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all mitochondria essentially retained

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its own translational machinery so the

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ribosome is still in there and there are

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certain critical components of oxidative

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phosphorylation system cycle cytochrome

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B cytochrome c k-- subunit 1 and 3 they

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still exist in every mitochondrial

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genome so mitochondrial ribosomes are

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required in order to and mitochondrial

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mitochondria from other species contain

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some additional variations of those

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genomes so my mitochondrial ribosomes

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are still required to do the translation

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within within mitochondria so and we

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have this quite unusual organism it's a

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eukaryotic organism called recliner

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Manas Americana and it appears that it

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contains a genome which is most

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bacterial alike genome among all known

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you can occur it mitochondrial species

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so this genome contains around 70 base

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pairs 70 70 kilo base pairs and it

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encodes for nearly 67 proteins so

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minimum number of proteins encoded by

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mitochondrial genome is four or five

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these this genome encodes

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67 proteins so and it's clearly by these

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genomic analysis Michael Gray had

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demonstrated back in 90s that the this

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mitochondria is closely related to an

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alpha Proteobacteria so and there were a

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number of genomic analyses done through

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a past couple of decades and kill very

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recently and this work was motivated and

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there gained its attention in the past

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few years when crystal structures of

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mitochondrial ribosomes became available

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from laboratories of Nanban and when

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Quinta Ramakrishnan so and through the

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rest of my talk I'm going to be talking

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about evolutionary processes that

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occurred within those mitochondrial my

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terrible mo structures and apparently

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they're quite different so we have

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fungal MITRE episome we have mammalian

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mightor episome and we have a parasitic

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mite ribosome and they all undergo

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substantial changes they accrete

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additional components most of them are

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proteins such that this is a this

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structure came out last year and it's

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very surprising because what happened is

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my tribe Azam nearly doubled its size

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compared to a bacterial ribosome and

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most of it it's just the proteins it's

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not a ribosome of RNA so and let me sort

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of start with maybe touch back on a

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question of mic travisano when he asked

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about the common core of bacteria it

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appears that there are some variations

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and some deletions with in bacterial

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common core and this is illustrated by

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this diagram secondary structure so even

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some bacteria do not have certain

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components and mostly salsa alpha

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alfa Proteobacteria so for example those

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regions mark in black here they do not

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exist or delete it or shortened even in

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alpha Proteobacteria so there is a

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remarkable resemblance between what one

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can see with ribosomal alpha

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Proteobacteria and the ribosome of

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requirements Americana which is a mighty

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ribosome we have the same deletions we

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have the same sequence conservation

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conservation essentially all those

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diagrams are mapped onto a secondary

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structure of E coli so and that was just

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the common core but now if we go and try

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to explore the entire array of available

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ribosome might arrive as almost

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structures there is a surprise that

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comes and essentially common core

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disappears so everything what we've

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talked about for for billions of years

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reduces down to an area which is

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highlighted in grey right here and the

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common core it uses down to just the

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origin that contains which contains the

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PTC a short region of exit tunnel and

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the platform of a large unit which

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interacts with a small subunit and there

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is a couple of other functional regions

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such as L 11 binding domain and the

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sarsen erasing loop so and what we did

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we reconstructed phylogeny based on

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structures of ribosomal RNA and I'm not

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going to go into details but just to say

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that we will use this phylogeny in order

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to time the events of what happens

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always mitochondria ribosomes and when

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we get this and use this phylogeny and

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try to map all different proteins among

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different species within different

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lineages

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so mitochondria retains a number of

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bacterial proteins so this mitochondria

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of reclaim honest Americana is the most

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bacterial like mitochondria it doesn't

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gain

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any additional proteins are little or no

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structural modification to that all the

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other ribosomes MITRE absorbs started

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creating additional proteins and those

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seven right here

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ml 41 43 45 46 49 53 and 54 appeared to

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be the oldest Oh nearly universal

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mitochondrial proteins so when I map

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those proteins onto three-dimensional

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structures

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it appears that those proteins all the

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Smita ribosomal proteins they are

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clustered centrally into two regions one

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is near the ready tunnel the second

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group is near central protuberance so

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let's start talking about the exit

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tunnel so we actually I mentioned

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earlier here that the Alpha

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proteobacteria ribosome and ribosome of

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requirements Americana contains some

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pretty heated regions which are marked

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in black here so and this but specific

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region here it's helix 16 helix 18 right

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here and it's shown based on a structure

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of e.coli right here of course there is

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no deletions but if you look what

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happened to or early protimeter ribosome

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from our reconstructions it appears that

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there are many early protocol and real

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proteins are clustered around this

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region so we hypothesize that those

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proteins are responsible for the build

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stabilization of this region and then in

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some later MITRE episome so if you go to

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a pista quante creates a fungi or

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mammalian ribosomes is helix completely

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disappears so that suggests that the

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existent secretion of those proteins

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just does a good job by itself

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RNA is no longer needed so basically

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we're just talking about a complete

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replacement and of of this region

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so now let's talk about the exit tunnel

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the tunnel also gets remodeled and it

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gets remodeled in two separate ways as

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especially when I'm talking about more

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advanced mitochondrial species I imply

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funda and a pistol quanta fungi and

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mammalian ribosomes this is what

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analysis was done and so mammalian

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ribosomes undergo into a mode of

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deletion so RNA gets shorter shorter and

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shorter and apparently we are getting

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more additional proteins in fungi

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ribosome the RNA doesn't get shorter but

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additional proteins get and stabilize

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this such that RNA itself undergoes

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refolding and it causes the change in

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the AG tunnel path

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so there is a new exit tunnel pass in

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East mitochondrial ribosome and it's

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diverted from its original path vertical

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path which exists in all other episodes

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including mitochondrial ribosomes

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bacterial ribosomes etc so and good so

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then a similar events occur within

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another region called central court

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appearance so we have accretion of those

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additional all these proteins ml 46 and

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ml 40 and even if George likes to talk

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about 5s RNA it is being lost upon

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accretion of those proteins probably

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because of their aesthetic issues and

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this coincides with the creation of

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protein BL 25 but then mitochondria gets

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protein ml 38 crediting into the

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basement of this central protuberance

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region and this is the reconstruction

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which corresponds to nearly loca last a

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pista quantum common ancestor of my

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the Condor ribosome and from this point

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the events took again two different

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approaches in mammalian ribosomes we the

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Galician continues on such that there is

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a deterioration of our RNA and

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mitochondria had to do a trick so it

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lacks its own RNA and it has to steal a

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tRNA molecule and just incorporate it

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into a central protuberance whereas in

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East mitochondria there are some

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additional Acree accretions expansions

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that associate with additional mighty

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ribosomal proteins and stabilize this

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region so again there are two different

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regimes in which those two mitochondria

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ribosomes can evolve so in order to

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conclude we we've shown that genetic

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corrosion has considerable effects on my

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mitre episomal structure and function

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loss of mitochondrial RNA stabilizes my

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tribus ohm and the structural fragility

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of metastable intermediates makes a more

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receptive to accommodating new elements

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patching occurs in an onion like fashion

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where new elements are created on to

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preexisting core and these structural

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patches are the addition of species

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specific pre-existing non-ribosomal

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macromolecules so all those new proteins

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they were available so now if I have a

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minute

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why all this is related to the origins

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of life we are talking about

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mitochondria here and I'm going back to

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the idea of the common core so the

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nature has managed to retain the common

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core over for billions of years the

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nature managed to contains a common core

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at the moment of at the moment of Luca

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when archaea split from bacteria and yet

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it just completely gave up on

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alpha product Iria got incorporated in a

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eukaryotic cell and it just under went

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through all this remodeling so what it

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suggests that there are very different

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types of evolutionary pressures that got

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applied onto this system of

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mitochondrial ribosomes and it just

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simply recorded all differences in

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evolutionary pressure and to me the

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critical difference is the difference in

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complexity the complexity of eukaryotic

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cell compared to complexity of bacteria

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when there is such a difference and it

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occurred over a short time it promoted

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such a severe changes in the structure

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of ribosome which suggests that when we

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got all different environmental changes

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changes in metabolism changes in cell

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composition upon split of Luca there

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were all of them we are not critical to

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the structure of the ribosome so it

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survived and maintained its own common

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very quick question you know I just have

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I have a question on a comment maybe the

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formulation of the question it would be

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to to ask truly why the ribosomal RNA in

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a carrots on in bacteria is so conserved

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in a sense because rather than looking

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at what's happening in the my to

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ribosome in fact I would say that it's

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really illustrating that the ribosomal

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RNA as you said indeed is under multiple

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order pressure but one that is becoming

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very apparent when you're looking at

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bacteria on a coyote is that it's

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completely remodeled the process of

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folding is completely controlled there's

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a huge amount of modification that takes

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place in a sequential fashion and in

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fact the ribosome sequence is conserved

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also because he has to switch well so I

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guess the short answer to your question

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would be when you need the car do you

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really care whether it's a Honda or camp

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or Toyota you don't but they're nearly

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identical they do the same functions and

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all the differences are just tiny

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details right right I'm sorry we're

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gonna have to interrupt this

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conversation we have another session