lecture on dna to proteins (the central dogma of molecular biology)

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michaelgray.org.uk

GENERAL PATHWAY PROKARYOTES vs EUKARYOTES

employees.csbsju.edu

Prokaryotes vs EukaryotesProkaryote Eukaryote

Site of DNA Replication Protoplasm Nucleus

Site of Transcription Protoplasm Nucleus

Site of RNA Splicing Rare Nucleus

Site of Translation Protoplasm Cytoplasm

Site of Recombination Protoplasm Nucleus

DNA REPLICATIONSEMI-CONSERVATIVE

STEPSUnwinding & Strand Separation

Helicase will unwind the parental strands breaks the H-bonds between bases

SSBP (Single-stranded DNA-binding proteins)

attaches to separated strands and keep them from annealing back

STEPS

STEPSUnwinding & Strand Separation

Topoisomerase breaks and rejoins the DNA double helix to relieve twisting forced by the opening of the helix (supercoils) mol-biol4masters.masters.grkraj.org

STEPSPriming

Primase catalyzes synthesis of RNA primers RNA primers attach to specific site of DNA strand (ORI region)

Synthesis of New Strands Leading strand (Continuous): DNA polymerase III catalyzes synthesis of new DNA strands at 5’ to 3’ direction (as single polymer) and extends the leading strand

STEPS

STEPSSynthesis of New Strands

Lagging strand (Discontinuous): DNA polymerase III extends the Okazaki fragments DNA polymerase I removes RNA primers and replaces it with DNA

Sealing the nick DNA Ligase forms covalent bonds from 3’ to 5’ direction to join the Okazaki fragments into a continuous strand

STEPS

DNA REPLICATION: Prokaryotespulpbits.com

DNA REPLICATION: Eukaryotes

For VIDEOS: https://www.youtube.com/watch?v=27TxKoFU2Nw

ENZYMES INVOLVED IN DNA REPLICATION

http://sydney.edu.au

TELOMERASE AND GENE EROSION

TELOMERES: ends of the DNA each chromosome is one continuous double stranded length of DNA with 2 telomeres

The 3’ to 5’ parent strand is copied no trouble as the leading strand BUT the lagging strand (parent 5’ to 3’) has small primers made every so often and filled in NOTE: no template at the end to bind to so when the primers are removed there is an overhang and exposes the DNA to digestion which would eventually eat into the genes at the end (gene erosion)

TELOMERASE AND GENE EROSION

TELOMERASE AND GENE EROSION

QUESTIONS????

REVERSE TRANSCRIPTION

REVERSE TRANSCRIPTION

OVERVIEW OF TRANSCRIPTION & TRANSLATION

TRANSCRIPTIONProcess of copying DNA to RNA

NOTE: only one strand of DNA, the template strand, is used to make mRNA

Does not need a primer to start and can involve multiple RNA polymerases Divided into 3 stages

Initiation Elongation Termination

For VIDEOS: https://www.youtube.com/watch?

v=WsofH466lqk

STEPS IN TRANSCRIPTION

STEPS IN TRANSCRIPTION

RNA POLYMERASERNA polymerase only transcribes one strand of the parental DNA molecule (the template strand) in the 3' to 5' direction

RNA polymerase will never add a T, instead it will add a U

Transcription ceases when RNA polymerase encounters special DNA sequences termed terminators that stop the process of transcription

RECALL…

INTRONS AND EXONSIntrons

sequences in the DNA that are NOT used to make mRNA or to make a protein

they are NOT transcribed

Exons

sequences in the DNA that are expressed or used to make mRNA and ultimately are used to make a protein

POST-TRANSCRIPTIONAL MODIFICATION IN EUKARYOTES

RNA SPLICING

REMOVAL OF INTRONS

5’ METHYLATED CAP AND POLYADENYLATION

PREVENTS DEGRADATION DURING TRANSIT FROM NUCLEUS TO CYTOPLASM

FINAL PRODUCT AFTER TRANSCRIPTION

TRANSLATION CAN BEGIN…www.can-con.ca

CONTROL DURING TRANSCRIPTION

PROKARYOTES

regulating genetic expression in nearby genes (e.g. LAC OPERON)

EUKARYOTES

involves the organization of chromatin and specific transcription factors which function by either changing the rate of transcription or altering the accessibility of chromatin to transcriptional machinery

the LAC OPERON

TRANSLATION: FROM VAGUE DNA CODES TO

MEANINGFUL TRAITS

blog.lionbridge.com

THE ASSEMBLY

barleyworld.org

For VIDEOS: https://www.youtube.com/watch?v=5bLEDd-PSTQ

IN SUMMARY…

EXAMPLE…DNA 5’-ATG GTT CGG TAG-3’ !

Replicating this strand.. 5’-CTA CCG AAC CAT-3’

mRNA transcript to undergo translation 5’-AUG GUU CGG UAG-3’

peptide from mRNA after translation Met-Val-Arg-STOP

THE GENETIC CODE

igoscience.com

NUCLEOTIDE CODES

A  

Adenine

G Guanine

C Cytosine

T Thymine

U Uracil

R Purine (A or G)

Y Pyrimidine (C or T)

N Any nucleotide

W Weak (A or T)

S Strong (G or C)

M Amino (A or C)

K Keto (G or T)

B Not A (G or C or T)

H Not G (A or C or T)

D Not C (A or G or T)

V Not T (A or G or C)

AMINO ACID CODES

Lys K Lysine

Met M Methionine

Phe F Phenylalanine

Pro P Proline

Ser S Serine

Thr T Threonine

Trp W Tryptophan

Tyr Y Tyrosine

Val V Valine

Asx B Asn or Asp

Glx Z Gln or Glu

Xle J Leu or Ile

Sec U Selenocysteine (UGA)

Pyl O Pyrrolysine (UAG)

Unk X Unknown

DNA SEQUENCE

NUCLEOTIDE SEQUENCE IS…

www.wiley.com

from the sequence… (AUA or AUG)-(CAU or CAC)-(GGU or GGC or GGA or GGG)-(AAU or AAC)-(CUU or CUC or CUA or CUG) !EXAMPLE: nucleotide sequence may be AUA-CAU-GGU-AAU-CUU !EXAMPLE: amino acid sequence is MHGNL or met-his-gly-asn-leu

LET’S HAVE A GROUPIE…

FORM YOUR GROUPSYou will be given set of questions and can discuss by group for the FINAL answer

The score of the group will be the score of each individual for today’s QUIZ

Submission is TOMORROW, 12noon, DB (Look for ate Malou)

READY???

QUESTIONSIn order to properly construct a protein, which process must occur first: transcription or translation? Why?

LAC operon: Under what conditions is the operator activated? Under what conditions is it inactivated?

Assume that the following sequence of bases represents a segment of DNA that is to undergo transcription. Determine the corresponding sequence of bases in the resulting RNA molecule.

3’-TAC GGC ATT GCA CAT TTT AGG GGC AAT ATT-5’

The following mRNA segment represents three codons. In each case determine what the anticodon would be on the appropriate tRNA.

AAA CGA AAU

SEE YOU ON MONDAY FOR YOUR EXAM 1

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