RNA Interference (RNAi): Mechanisms and Components - Prof. Vitiello, Study notes of Molecular biology

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Not (^) knockouta Reduce (^) Slide 3 of RNAi or knockdown viruses, Silence gene expression, transposable elements

long non-coding RNA

(microRNAlendogenus i Processes

Into Short/small guided RNA

miRNA (^) SiRNA (^) exogshort interferingd miRNA SiRNA knockdown specificfortransposoa short hairpin

• bunch or 7 homosaping, C

A Different part of genome

or invading RNA^ or^ DNA

trying to^ incorporate^ itself

transposable elements^ into the cellular (^) genome

in place its^ not suppose

to be^ invading nucleotides.

• You don't have introns in mature RNA

RdRP (^) or (^) RNA dependent RNA (^) polymerase^ exogenous DICER RNA-dependent RNoMymeraze MRNA

80s URNA 60s

40s (^) 18s anticodon-codon ItS So 235 285 (^285) Peptidyl transferase

185 IS^ tRNA

STRUCTURE

MENA CRNA

codon

codon on the (^) MENA that (^) determines the (^) proper amino acid codon on the meny is^ going to tell thatdend^ it^ needs^ to^ be

never

Rilotyeme

zero S^ zero^ -

#When we (^) say the run (^) occurs With (^) Zero

proteins, what^ we^ are^

meaning is that catalytic domain^ ,^ where^ the^ action^ or Removing the^ protein^ the^ rxn^ will^ still^ occur

active site is there are not proteins.^ ·^ no^ amino^ acids^ in^ active^ site

*The enzyme is not a protein.The ·^ When^ you^ Isolate^ 28s^ rRNA^ and^ test^ its^ function

enzyme is^ RNA^ , ribose

• the behaves (^) as a (^) ribozyme (peptidasferase

T

should be amino^ acid

F

after

attacheshere T e

F

should be RWAs P Site

F NostartsWeSee

appears

gi RNase P

tRNA STRUCTURE ↑ attacks amino (^) acyl tRNA^ Synthetase adenylylation (^) onosphate

ATP X

adenosine zi

charging

high

• the^ energy we^ use^ when^ we^ are^ doing peptidyl transferase^ rxn^ to^ form^ that

peptide bond.

GTP-elF elf) elFA elfE kinds to smallSubunit^ & to prevent (^) bindste

large subunit

tohind

& elin e helia se

unwinds

brings -

met-tRNAjmet

to the psite

Poly i

binding protein

noncoviently

Interact (^) with

Poly A^ binding

protein witail^ associates

Accommodation

* where^ tRNA^ turns^ , so the^ amino^ acid^ on that^ nation

tRNA and the peptide chain are

bascially!

facing each^ other^ so^ that^ they can^ react^ - y

peptidylnsferase

EF-TU

Factor (^) binding center ref- tv

Translocation

accommodation occured START OF^ TUN Accommodation RXN already peptidyl occurred?^ transferase nVC attack on peptidyl aming acid Moves in S'to 31 transferase in (^) p (^) site direction by one RXN (^) ↑ codon Site A^ Site now Ribosome

is in^ a^ hydride^ - A^ site

state (^) peptidona emptyready

*hybrid state means for

when a^ peptide^ bond^ next^

URNA

forms and (^) polypeptide

andRNA^ in^ the^ A^ Site

• now needs translocation to occur.

USCAR MEND Oscar mRNA

IRP WIRON

Cup to binds Bruno

* where our cup protein binds to^ the^ Bruno

factor and that cut proteins 4E-BP

domain will in^ turn^ kind will^ bind

Initiation factor^ IE.

Basically prevents^ ↑G from (^) binding

and leads from^ the others^ not^ to

bind. preventing from^ one (^) binding , you (^) prevent the (^) rest from (^) binding and

translation will not occur

* IRP^ WIO^ IRON^ will bind to^ the IRp can

the (^) regulatory element IRE^ Hind^10 X

and that^ prevents our

Initiation factors^ from^ M function and^ further further (^) Binding

* IRP^ W/^ IRON^ the^ IRP^ will^ IRP

hind IRON^ at^ fEz^ and

that means^ it^ can't^

&IRP can

NOX HIND (^) TO IRE hind the^ regulatory

element.

means that the^ IRP^ can^ bind^ either^ Iron^ or^ the^ IRE^ , it^ cannot^ do^ both!

• upstream open (^) reading Frame (^) , (^) basically cause tin to initiate (^) and (^) terminate (^) very quickly

in sequence.

That makes^ tribosome^ dissociate^ really (^) quickly , and (^) not (^) to rehind. The (^) actual gene behind

them won't^ get translated

• GCNY^ , particularly about (^) the difference in^ level^ of^ amino^ acid^ , when GCNT^ is involved in amino (^) acid (^) biosynthesis (process^ in^ body where we make amino^ acid)

when we^ have^ amino^ acid we^ don't^ need^ that^ to turn on.

• VORF^ is^ being translated^ , (^) making Ribosome^ skip^ over^ GCNY^ transcript part my causing theribosome^ to^ quickly dissociate^ , but^ when^ we^ have^ low^ or^ no^ amino acids we^ want^ the^ GCNT^ to be (^) translated. De to (^) low levels =^ GTP (^) hydroluce more (^) slowly