November 20, 2013

Molecular Genetics: RNA

and Transcription
Reading: Chapter 15
 Last time
Chargaffs Rule: A = T and G = C
Bases are paired so the two DNA strands are
complementary
DNA reproduces by semiconservative replication;
each strand is copied to make new complementary
strands
The leading strand is copied continuously (3 to 5);
the lagging strand is copied discontinuously in
segments (Okazaki fragments)
DNA polymerase III extends the DNA strand and
needs RNA primers to begin
The replisome organizes many of the enzymes
needed for DNA replication
Telomerase extends the lagging strands at the
ends of each chromosome
Some People Cant Have Nutrasweet
 Phenylketonuria (PKU)
Phenylketonurics cannot metabolize
phenylalanine, so phenylalanine and
phenylketones accumulate and appear in the
urine
Aspartame (Nutrasweet) is composed of two
amino acids, phenylalanine and aspartic acid
A child with undetected PKU may have a smaller
than normal head, epilepsy and mental
retardation
http://www.ygyh.org/pku/have08.htm
 Phenylketonuria Is Inherited
PKU is a recessive genetic disorder
occurring in 1 in 12,000 human infants
It is caused by a defective gene for the
enzyme phenylalanine hydroxylase
One Gene-One Polypeptide Hypothesis

A gene specifies the structure of a protein

(or more precisely a single polypeptide)
This hypothesis was tested by George
Beadle and Edward Tatum using the
bread mold Neurospora
There are important exceptions to the
hypothesis
One Gene-One Polypeptide

What Beadle and Tatum

showed:
Yeast mutants that require
arginine in the medium
have a defective enzyme in
the pathway for arginine
synthesis
 The Central Dogma
Information flow goes from DNA to RNA to protein
DNA RNA Protein

Transcription: DNA RNA

Translation: RNA Protein
Transcription
A segment of DNA is
copied to make a
complementary strand
of mRNA
 Transcription and
Translation in Prokaryotes
mRNA is
transcribed
off of DNA
Ribosomes
on the mRNA
are
translating
the mRNA to
make protein
Transcription
A segment of DNA is
copied to make a
complementary
strand of mRNA
 Transcription
Coding strand
= sense
strand
Template
strand =
antisense
strand

Upstream Downstream
 Direction of RNA Synthesis
The new RNA is assembled 5 to 3
As with DNA, the nucleoside triphosphate
can only be added to the 3 end
 Stages in Transcription
Initiation RNA Polymerase binds to the
promoter and starts transcription a little
downstream from there
Elongation RNA polymerase copies the
template strand from the 3 end toward the 5
end (the same as DNA polymerase)
Termination Occurs when RNA polymerase
recognizes a stop sequence
Transcription in Prokaryotes
RNA is
transcribed
from DNA by
RNA
polymerase
The subunit
is required to
recognize the
start site
Transcription in Prokaryotes

RNA polymerase recognizes a start signal

at nucleotide -35
DNA unwinds beginning at nucleotide -10
RNA transcription starts at nucleotide +1
Transcription in Prokaryotes
As RNA is transcribed,
the DNA unwinds at
the downstream end of
the bubble and
rewinds at the
upstream end of the
bubble
RNA polymerase
moves along the DNA
until it reaches the
termination point
 Kinds of RNA
mRNA (messenger RNA)
rRNA (ribosomal RNA)
tRNA (transfer RNA)
snRNA (small nuclear RNA)
SRP RNA (signal recognition particle)
Small RNAs: miRNA (micro RNA) and
siRNA (small interfering RNA)
 Transcription in Eukaryotes

RNA polymerase I makes rRNA

RNA polymerase II makes mRNA
RNA polymerase III makes tRNA
 Prokaryotes vs. Eukaryotes
In prokaryotes, mRNA is translated to
make proteins right away
In eukaryotes, mRNA must leave the
nucleus because proteins are made in the
cytoplasm
In eukaryotes, mRNA is modified in
important ways
 Transcription in Eukaryotes

Transcription factors are involved in

recognizing the initiation site and recruiting
RNA polymerase II to the DNA
 Post-transcriptional
Modification in Eukaryotes
5 Cap: A methylated GTP is added to the 5 end
of the RNA. This reduces degradation from the 5
end (the ribose is hooked on backwards)
 Post-transcriptional
Modification in Eukaryotes
PolyA tail: A series of adenine nucleotides
(100-200) are added to the 3 end of mRNA by
polyA polymerase
 Post-transcriptional
Modification in Eukaryotes
Splicing: Introns
(noncoding
regions) are
removed from
mRNA by splicing

DNA-mRNA hybrid
shows DNA loops
where mRNA is
missing introns
Removing Introns in Eukaryotes
Introns are removed in the nucleus
The spliceosome removes introns
snRNA in small nuclear ribonucleoprotein particles
(snRNPs) recognizes intron/exon junctions
 Alternative Splicing
mRNA can be spliced in different ways in
different tissues
Because of alternative splicing, a single
gene can actually produce more than one
polypeptide product
Alternative splicing allows the human
genome to make 120,000 or so proteins
using only 25,000 or so genes
One gene one (family of) polypeptide
 Summary
One gene specifies one (family of) polypeptide
The Central Dogma says that information is
transferred from DNA to RNA to protein
Transcription is the process by which mRNA is
made as a complementary strand to the template
strand of DNA
In eukaryotes, mRNA must exit from the nucleus
after posttranscriptional modification including
addition of a 5 cap of methylated GTP, the addition
of a 3 poly-A tail, and the removal of introns
Removal of different sets of introns by alternative
splicing enables a single gene to code for more than
one polypeptide
mRNA exported from the nucleus
does not contain

A) polyA tail
B) 5 cap
C) exons
D) introns
RNA polymerase starts transcribing DNA at
the start site and moves

A) downstream on the coding strand

B) upstream on the coding strand
C) downstream on the template strand
D) upstream on the template strand
If RNA has the sequence
5 A U G U C A C 3
the template strand of DNA has the
sequence
A) 5 U A C A G U G 3
B) 5 T A C A G T G 3
C) 3 T A C A G T G 5
D) 3 U A C A G U G 5
E) 3 A T G T C A C 5
 Next Time

Molecular Genetics:
The Genetic Code

Reading: Chapter 15
 Coming Up
Nov 22, 25 (Fri, Mon)
Protein Synthesis Chapter 15
Nov 27 and 29 Thanksgiving Holiday
Dec 2, 4 Gene Expression Chapter 16
and SET
Dec 6, 9 Development - Chapter 19
Dec. 12 (Thursday)
Exam IV - 10:40 a.m.
Makeup Exam -11:50 am