GENE EXPRESSION

Process by which information from a gene is used in the synthesis of a functional gene product

GENE EXPRESSION

Gene products include; proteins, rRNA, tRNA or SnRNA genes Gene expression is virtually the same in all eukaryotes Differences with prokaryotes

 Synthesis of RNA from a DNA template catalyzed by RNA polymerase

RNA polymerase is of 3 types; RNA pol 1 - rRNA RNA pol 11 - mRNA RNA pol 111 tRNA Has 3 recognizable steps; Initiation Elongation Termination

TRANSCRIPTION IN EUKARYOTES

 Involves recognition of the start point upstream of the gene

Determined by a promoter sequence in addition to enhancers & Transcription factors (TF) Promoter seq; a unidirectional sequence that instructs the RNA pol where to start synthesis & in which direction synthesis should continue Most common promoter is TATA box, -30 base pairs from the start site of transcription Assembly of TF & RNA pol to the promoter form a transcription initiation complex known as an Open Promoter Complex.

Initiation

 Non coding strand acts as a template for RNA synthesis

No need for primers RNA pol uses base pairing complimentarily to create an RNA Copying is in the 3- 5 direction

Enlogation

Elongation also involves a proofreading mechanism that can replace incorrectly incorporated bases

Termination
No strong termination sequences like prokaryotes

RNA polymerase II continues transcribing up to 1000 to 2000 nucleotides beyond where the 3' end of the mature mRNA will be.

Eukaryotic translation

Cytoplasm

DNA

Transcription
RNA

RNA Processing
mRNA G
AAAAAA

AAAAAA

Export
Nucleus

Capping of the RNA Occurs at the 5' end A methylated guanine nucleotide is added Cap is for recognition of the mRNA by ribosomes during translation
Polyadenylation (AAUAAA) Involves addition of a string of adenine nucleotides to the 3' end AAUAAA is recognized by the appropriate enzymes which cleave the RNA 10 to 30 nucleotides downstream & a string of adenine nucleotides Protect the RNA from degradation & plays other regulatory roles

GENE PROCESSING OR EDITING

 Introns in some RNAs can self-splicing

Intron removal

No enzyme is required(enzyme activity is within the intron itself), such RNA enzymes are termed ribozymes Class II RNAs do require enzymes to remove their introns Splicing is carried out by spliceosomes Spliceosomes able to detect intron/exon boundaries, cleave the RNA at the appropriate point & join adjacent exons together to produce the mature mRNA

 Animal genes are large while plant genes tend to be much smaller (1-2 Kb) Animal genes have numerous large introns while plants have fewer and smaller introns Plant transcripts retain introns more often than do animal transcripts (30% of all genes in the model plant, Arabidopsis, compared to 10% in humans)

Differences between transcription in plants and animals

 Occurs in the cytoplasm

DIFFERENCES WITH PROKARYOTES

RNA pol is made up of 5 subunits; 2, , 1 & Sigma subunit recognise the promoter seq Termination involves formation of the hair pin loop structure Release of the mRNA is either dependent or independent on the rho protein No modification of the new transcript

 Conversion of the information of mRNA into proteins

mRNA produced by transcription is decoded by the ribosome to produce a specific amino acid chain Occurs in the ribosomes Ribosome facilitates decoding by inducing the binding of tRNAs with complementary anticodon sequences to that of the mRNA The tRNAs carry specific amino acids that are chained together into a polypeptide as the mRNA passes through

TRANSLATION IN EUKARYOTES

Activation
The correct amino acid is covalently bonded to the correct tRNA The amino acid is joined by its COOH to the 3' OH of the tRNA by a peptide bond Charged tRNA has an amino acid linked to it

Structure of tRNA

Clover shaped
Has various arms
Aligns each amino acid with the corresponding codon 3 end has the 5- CCA sequence to which aa are linked The opposite end contains the anticodon loop Contains modified bases

 Requires hydrolysis of ATP and GTP

Initiation

Involves the interaction of proteins with a special tag bound to the 5' cap for recognition Results in formation of a complex containing the mRNA, the ribosome and the initiator Met-tR

The initiation complex bound to the 5 cap structure scans in a 5 to 3 direction until initiating AUG is encountered
Initiation complex binds upstream of initiation codon

Enlogation

Termination Translation is terminated at one of three stop codons (UAA, UAG & UGA).

Termination codon at the A site is recognized by the release factor instead of a tRNA
The release factor binds the termination codon The peptide chain is then released followed by dissociation of the tRNA and the ribosome

 Translation begins before transcription is complete

Differences in prokaryotes

Translation in prokaryotes is polycistronic

No post translation modifications

POSTTRANSLATIONAL MODIFICATION IN EUKARYOTES

Involves chemical modification of a protein after translation Can be done through changing the chemical nature of an amino acid or structural changes Amino acids can be removed from the amino end of the protein, or cut the peptide chain in the middle This extends the range of functions through attachment to other biochemical functional groups e.g acetate, phosphate, various lipids & carbohydrates,

 Consists of 64 triplets of nucleotides called codons Codon; a seq of 3 bases that specify an amino acid, a start or stop signal

GENETIC CODE

Code word Colinearity Start codon (AUG) Stop signal(UAA, UAG, UGA) Degeneracy Universality

Xtics of the Genetic Codon

CODON BIAS Except 2 of the amino acids (Met and Trp) can be encoded by from 2 to 6 diff codons However certain codons are preferred over others. In man, alanine is encoded by GCC four times as often as by GCG This reflects great translation efficiency by the translation apparatus for certain codons over their synonyms

Regulation of gene expression

Refers to the control of the amnt & timing of appearance of the functional product of a gene Vital as it allows a cell to produce only the gene prdts it requires This gives the cell control over its structure & function thus basis for cellular differentiation & morphogenesis

Terms used to describe genes

House keeping gene Is a gene that is transcribed continually also called constitutive gene, eg actin

Facultative gene Is a gene which is only transcribed when needed

Inducible gene Is a gene whose expression is either responsive to environmental change or dependent on the position in the cell cycle.

Regulation of gene expression

Control occurs on many levels of gene expression

Controls the no. of mRNA transcribed

Regulation of transcription

Modulators of transcription; Activators -enhance the interaction between RNA pol & a promoter Enhancers -sites on the DNA helix that are bound to by activators so as to loop the DNA bringing a specific promoter to the initiation complex Specificity factors - alter the specificity of RNA pol making it more or less likely to bind e.g sigma factor

Repressors (silencers) - bind to non-coding sequences to the promoter region impeding RNA pol progress along the strand,
General transcription factors -position RNA pol at the start of a start site & then release it to transcribe the mRNA

 Addition of a cap and tail to the RNA identifies the mRNA by the ribosomes & prevents degradation Splicing can determine whether mRNA gets translated, If not processed it is not transported out of the nucleus thus not translated
Exon shuffling where some exons are exchanged changing the protein produced

Post transcription regulation

Translation Control
Controlling the no. of ribosomes allowed to attach a single mRNA Controlling the rate at which each ribosome transcribes a message. Use of inhibitory proteins that prevent the translation of mRNA

Regulation of RNA Longevity

Lifespan of mRNA determines no. of times it can be used to create proteins Eukaryotic mRNA more stable than for prokaryotes AUUUA is a signal for early degradation

SIGNAL TRANSLATION
Purpose; Regulate cell growth & differentation Co-ordinate diff phjsiological processes Maintain homoestasis

Forms of translation Use of molecular messengers (hormones & paracines) Use of non-diffusible cell adhesion proteins

Molecular messengers
Hormones Are molecules released by endocrine glands into circulation to act on specific target cells widely distributed 3 categories; steroids, Proteinaceous & amino acid related Steroids; Insoluble bt can freely cross the plasma membrane Binds to specific recptors forming a complex Complex binds to speciic regions on DNA to regulate transcription

Proteinaceous & aminoacid related

Are hydrophillic Cannot cross the plasma membrane thus interact with surface receptors Induces a 2o messenger which results in reguation of transcription Paracrines Hv local action & restricted to neighboring cells 4 categories; mitogens, trophic factors, chemoattractants & pleiotrophic Can be hydrophilic or hydrophobic

Non-diffusible adhesion molecules

Activate physical cell to cell matrix interaction regulating gene transcription

Signal Translation in Plants

Plants have evolved different signaling mechanisms as their hormones function differently from in animals

Plants also cope with environmental changes differently as they dont physically escape except possibly through reproduction.