Gentica

Esto es un gen

HealthCare

Composicin, estructura y funcin del ARN

Secuencia transcrita (ARNm)

GCGCG..TATATACCAA..ATGTGCTTCTTACAGGAAT..TAACGCGGG..AATAAAGCGCCTTT..
Amplificador Promotor Represor Exon Intron Secuencia de finalizacin

Secuencia codificante

Composicin del ARN

Estructura del ARN

Estructuralmente una molcula de ARN es parecida a una molcula de ADN, con las siguientes diferencias:

1. Bases nitrogenadas: purcas [adenina (A) y guanina (G)] y pirimdicas [citosina (C) y Uracilo (U)]. 2. Ribosa (azcar de 5 carbonos) 3. Grupo fosfato (acido fosfrico)

1. El azcar del ARN es una ribosa y no una de desoxirribosa. 2. En el ARN el Uracilo remplaza a la Timina, como base nitrogenada. complementaria de la Adenina. 3. El ARN es de hebra simple. 4. El ARN es mucho ms pequeo que el ADN.

ARNt

Transcripcin en eucariotas (sntesis del ARN)

Principales elementos implicados

Transcripcin en procariotas

1. 2. 3. 4. 5.

ADN conteniendo el gen a ser transcrito. Factor sigma (protena) ARN polimerasa (una sola) dNTPs: dATP, dUTP, dCTP, dGTP Factor Rhro (proteina) - opcional

ARN Polymerasa (aloenzima compuesta de varias sibunidades )(Mg++)

Etapas de la trascripcin

a) Iniciacin b) Elongacin c) Terminacin

factor sigma

RNA polymerase lacks the exonuclease activities. As a result, RNA polymerase does not proof-read and is much more error prone than DNA polymerase. Accuracy: ~ 1 error in 105 bases

1. Iniciacin
Promotor procariota

E. coli RNA polymerase + (factor sigma)

spacer The -35 and -10 boxes contain consensus sequences

Strands melted from 10 to about +1 (TIS) producing a single stranded DNA in the active site of RNA Polymerase.

2. Elongacin Elementos adicionales

No necesita primers! Una vez que se han aadido varios nucletidos, el factor sigma se disocia y la elongacin continua.

En E. coli, 50 nucletidos/segundo a 37 grados C Los codones stop no detienen la ARN polimerasa..

3. Finalizacin
A) Terminacin directa

Repeticin inversa rica en G:C y residuos de U sirven para soltar la ARN polimerasa. Esta secuencia forma una horquilla

B) Terminacin dependiente del factor Rho (Helicasa dependiente del ATP)

Sitio rico en G/C

Transcripcin en eucariotas
Pausa de la ARN Pol. El factor Rho alcanza el complejo

Se separa el complejo de elongacin

Principales elementos implicados

1. 2. 3. 4. 5. ADN conteniendo el gen a ser transcrito. Protena que interacta con la caja TATA (TPB: TATA binding protein). Factores de transcripcin (TF: trancription factors) TFIID, TFIIB, TFIIA, TFIIF, TFIIE, TFIIH. ARN polimerasa (I, II y III). La ARN polimerasa I transcribe los ARNr, la ARN polimerasa II transcribe los ARNm y la ARN polimerasa III transcribe los pequeos ARNs (p.e. ARNt). La ARN polimerasa II contiene un dominio llamado CTD (carboxy terminal domain) compuesto de 52 repeticiones del hepta-pptido YSPTSPS) que es esencial para su actividad. dNTPs: dATP, dUTP, dCTP, dGTP Amplificadores de la transcripcin (enhancers) tales como el ARC (activator recruited complex), CBP (CREP binding protein), DRIP (vitamin D receptor interaction protein), SP1 (specificity protein 1) SREBP (sterol response element binding protein), etc. Enzimas enfundadoras 5 (5 capping enzymes): ARN trifosfatasa, guanililtransferasa, ARN metiltransferasa. Factor de ruptura y poliadenilacion (CPSF: Cleavage and Polyadenylation Specificity Factor) Factor estimulador de ruptura (CstF: Cleavage Stimulation Factor) Poliadenilasa polimersa o Poli A polimerasa (PAP: Polyadenylate Polymerase) Protena nuclear protectora de la cola Poli-A (PABPN1: Nuclear Polyadenylate Binding Protein). Espliceosoma (complejo ribonucleoproteico)

RNAs polimerasas en eucariotas

6. 7.

8. 9. 10. 11. 12. 13.

1. Iniciacin (ARNm)

Amplificadores

TBP

Sitio de iniciacin

Plants have AGGA instead of CAAT!!

2. Elongacin

pol II CPSF CstF Enzimas enfundadoras 5 1. ARN trifosfatasa 2. guanililtransferasa 3. ARN metiltransferasa
Cleavage and Polyadenylation Specificity Factor Cleavage Stimulation Factor

5 methylguanosine cap added, prior to transport out of nucleus.

3. Finalizacin

4. Modificacines post-transcripcionales

Cleavage and Polyadenylation Specificity Factor

CPSF

pol II

Cleavage Stimulation Factor

CstF
pol II

35b

PABPN1
Nuclear Polyadenylate Binding Protein

poly a tail ( several to 250) added after cap; degredation rapid if tail missing

Modificacines post-transcripcional
Small nuclear RNA (snRNA) Plays structural and catalytic roles in spliceosomes, the complexes of proteins and RNA that splice pre-mRNA in the eukaryotic nucleus.

ARN maduro

Small nuclear RNA (snRNA)

small nuclear RiboNucleoProteins

UAA UAG UGA

Enhancers
Enhancers are DNA regions which are usually rich in transcription factor binding sites and/or repeats. They enhance transcription of the responsive promoter independent of orientation and position. Some transcription factors ("Enhancer-binding protein") bind to regions of DNA that are thousands of base pairs away from the gene they control. Binding increases the rate of transcription of the gene. Enhancers can be located upstream, downstream, or even within the gene they control

Prokaryotic vs. Eukaryotic Transcription

Prokaryote All promoters upstream of functional gene Main promoter consensus sequences TATAAT (-10) and TTGACA (-35) One RNA polymerase with subunit makes mRNA, tRNA, rRNA No enhancers mRNA is primary transcript ready to go short lifetime (just a few minutes)

Eukaryote Promoter positions differ for each polymerase- not all upstream Main consensus sequence TATA box (-25) and CAAT box (-60 to -120) Plants have AGGA instead of CAAT RNA POL I rRNA RNA POL II mRNA RNA POL III ss rRNA, tRNA DNA enhancer regions work with some promoters to increase transcription Initial product of transcription is not usable mRNA. Primary transcript must be processed to form mRNA. Longer lifetime (hours/days)

Transcriptasa inversa

Initially discovered by Temin and Baltimore copies RNA into DNA. The HIV enzyme is a dimer of a p66 and a p51 cleaved subunit. The two protein differ in a region of the crystal structure wich covers the RNase H region (p51 lacks the activity). The crystal structure reveals resemblance to T7 pol and KF.

Example Human Immunodeficiency Virus (HIV)

The flow of genetic information 1960

DNA
transcription

The flow of genetic information 1982

DNA
reverse transcription t r a n s c r i p t i o n

mRNA
translation

hnRNA
splicing

tRNA

rRNA

ribozymes

Protein

Molecules with effector function

mRNA
translation

Protein

Molecules with effector function

The flow of genetic information 2004

DNA
reverse t transcription r a n s c r i p t i o n

hnRNA
s p l i c i n g

tRNA

rRNA

ribozymes

mRNA
translation

Other noncoding RNAs ncRNAs Molecules with effector function

Protein