10/25/2011

The first stage in the flow of information from DNA to polypeptide is the transcription of a DNA nucleotide sequence into an RNA nucleotide sequence.

10/25/2011

RNA polymerase

In transcription, the DNA helix unzips

RNA nucleotides line up along one strand of the DNA following the basepairing rules The single-stranded messenger RNA peels away and the DNA strands rejoin

DNA of gene

Promoter DNA Initiation

Terminator DNA

Elongation

Termination

Growing RNA

Completed RNA

RNA polymerase

Three main kinds of RNA molecules are transcribed:

messenger RNA transfer RNA ribosomal RNA

10/25/2011

Messenger RNA (mRNA)

is a single strand of RNA that carries the information for making a protein.

Transfer RNAs (tRNAs)

is a single strand of RNA that folds back on itself to form a specific shape.

Ribosomal RNA (rRNA)

is in a globular form, is an important part of the structure of ribosomes and has catalytic functions needed during protein synthesis.

Most RNA is synthesized by one of three RNA polymerases, enzymes present in all cells. The three RNA polymerases differ in the kinds of RNA they synthesize.

10/25/2011

RNA polymerase I
catalyzes the synthesis of several kinds of

rRNA molecules that are components of the ribosome;

RNA polymerase II
catalyzes the production of the protein-

coding mRNA;

RNA polymerase III

catalyzes the synthesis of tRNA and one of

the rRNA molecules.

10/25/2011

Promoter :

a region of DNA where RNA polymerase binds to initiate transcription

Promoters contain specific, highly-conserved sequences of DNA Transcription factors contain DNA-binding domains that recognize these specific sequences Common promoter elements are shown below; many eukaryotic genes have these (though not all) TATA box is important

10/25/2011

A characteristic feature of a transcriptional terminator is the presence of a short sequence that is complementary to the sequence just preceding it. and so can anneal together resulting in the formation of a stem-loop structure. In most terminator sequences, the stemloop structure is followed by a run of U residues.

10/25/2011

10/25/2011

10/25/2011

1. 2. 3. 4. 5. 6.

Pre-mRNA containing introns and exons 5 end of premRNA capped with modified nucleotide Poly-A tail added to 3 end Introns removed and exons spliced together Mature mRNA transported into cytosol Translation at ribosome

Each end of a pre-mRNA molecule is modified in a particular way

The 5 end receives a modified nucleotide cap The 3 end gets a poly-A tail

A modified guanine nucleotide added to the 5 end

TRANSCRIPTION DNA Pre-mRNA RNA PROCESSING mRNA Ribosome TRANSLATION

50 to 250 adenine nucleotides added to the 3 end Polyadenylation signal 3 AAAAAA AAUAAA 3 UTR Poly-A tail

5 G P P P 5 Cap
Polypeptide

Protein-coding segment

5 UTR

Start codon Stop codon

Figure 17.9

10/25/2011

RNA splicing
Removes introns and joins exons
5 Exon Intron Pre-mRNA 5 Cap 30 1 31 Exon 104 Intron 105 Exon 3 Poly-A tail 146 Introns cut out and exons spliced together Poly-A tail 146 3 UTR

TRANSCRIPTION

DNA Pre-mRNA

RNA PROCESSING mRNA

Ribosome TRANSLATION

Coding segment mRNA 5 Cap 1 3 UTR

Polypeptide

Is carried out by spliceosomes in some cases

5 Protein
1

RNA transcript (pre-mRNA) Exon 1 Intron Exon 2

snRNA snRNPs

Other proteins

Spliceosome

Spliceosome components mRNA Exon 1 Exon 2 Cut-out intron

10

10/25/2011

The presence of introns

Allows for alternative RNA splicing

Ribozymes
Are catalytic RNA molecules that function as

enzymes and can splice RNA

Proteins often have a modular architecture

Consisting of discrete structural and functional

regions called domains

In many cases
Different exons code for the different domains

in a protein
DNA Exon 1 Intron Transcription RNA processing Translation Domain 3 Gene Exon 2 Intron Exon 3

Domain 2 Domain 1

Polypeptide

11

10/25/2011

The mRNA has a non-coding leader sequence at its 5 end. The leader has recognition sites for ribosome binding, which properly position the ribosomes to translate the message.

12

10/25/2011

The start codon follows the leader sequence and signals the beginning of the coding sequence that contains the actual message for the polypeptide At the end of each coding sequence, a stop codon signals the end of the protein

The stop codonsUAA, UGA, and UAGend both bacterial and eukaryotic messages. They are followed by non-coding 3 trailing sequences, which vary in length.

13

10/25/2011

Following transcription, the transcribed information in the mRNA is used to specify the amino acid sequence of a polypeptide This process is called translation because it involves conversion of the nucleic acid language in the mRNA molecule into the amino acid language of protein.

14

10/25/2011

A specific enzyme called an aminoacyltRNA synthetase

Joins each amino acid to the correct tRNA

Ribosome Structure
Ribosomes are complexes of RNAs and proteins.

15

10/25/2011

Ribosomal RNA
Ribosomal RNA (rRNA) is single stranded RNA that folds into a complex structure due to hydrogen bonding. The peptidyl transferase activity of the ribosome appears to be associated with the RNA in the ribosome rather than the proteins.

Ribosome Binding Site

The Ribosome Binding Site (RBS) or Shine-Dalgarno sequence (which was characterized by John Shine and Lynn Dalgarno) is a short sequence that base pairs with the 16S rRNA. The RBS will be near the start codon of prokaryotic genes.

16

10/25/2011

The process of protein synthesis has three distinct stages:

initiation, repeating cycles of elongation, termination.

Initiation of protein synthesis is essentially the same in all organisms

In Escherichia coli, the ribosome binding site has the consensus sequence 5 AGGAGGU-3 and is located between 3 and 10 nucleotides upstream of the initiation codon. Shine Dalgarno sequence: The ribosome binding site upstream of an E. coli gene

17

10/25/2011

Initiation factors bind to the small ribosomal subunit, which then binds to mRNA in the region of AUG, the start codon. The initiator tRNA binds to the start codon, followed by binding of the large ribosomal subunit.

18

10/25/2011

Elongation is a cyclic process in which amino acids are added one by one to the growing polypeptide chain. Elongation proceeds in the 5 3 direction along the mRNA. The polypeptide chain grows from its amino end to its carboxyl end.

19

10/25/2011

Termination, the final stage of translation, occurs when the ribosome reaches one of three stop codons. The A site binds to a release factor, which triggers the release of the completed polypeptide chain and dissociation of the translation complex.

20

10/25/2011

An overview of transcription and translation

21

10/25/2011

The genetic code was assumed to be universal, meaning that each codon specifies the same amino acid in all organisms, with a few exceptions.

22

10/25/2011

A gene
Is a region of DNA whose final product is either

a polypeptide or an RNA molecule

23