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Genetic Organisation. In Prokaryotes. Transcrition and translation occurs in same compartment (cytoplasm) Simultaneous; m-RNAs are short-lived (afew minutes) No splicing M-RNA is not polyadenylated No IRES in eukaryotes No introns in prokayotes (except some bacteriophages). 5’-3’ direction
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In Prokaryotes • Transcrition and translation occurs in same compartment (cytoplasm) • Simultaneous; m-RNAs are short-lived (afew minutes) • No splicing • M-RNA is not polyadenylated • No IRES in eukaryotes • No introns in prokayotes (except some bacteriophages)
5’-3’ direction Any strand of DNA can be transcribed No need for helicases, topoisomerases, primers RNA polymerase: 4 chains 2alpha, beta, Beta’ Promoter is recognised by the factor sigma Transcription
In some cases termination rho dependent factors, which are helicases • Rho utilisation site 80-100 bp upstream of actual terminator • In E. Coli other factors: tau, nus
70 s = 50 S and 30 S 50 S subunits: 23 S and 5 S RNA molecules 30 S subunits: 16 S RNA and 21 polypeptides rRNA binds to m-RNA at spesific sequences: Shine-Dalgerno sequence (RBS) partly complementary to the 3’ end of 16 S RNA In bacteria ribosomes
Secretion of the proteins • Many proteins exert their functions on the cell surface or in extracellular environement they should across the cytoplasmic membrane • GSP Sec dependent pathway • Proteins utlising GSP have a specific sequence at their N termini, which is cleaved during the transport
In Gram positive bacteria GSP is sufficient but ın Gram negative bacteria Proteins reach only to the periplasmic space
Gram negatives have addional mechanism: Sec dependent and Sec independent Sec dependent systems • Type II secretion system: A multiprotein complex transports proteins from the periplasmic space to the outside • Type V secretion system: The proteins have an additional sequence at the C terminus, forming pores in the outer membrane (aototransporters)
Sec independent systems • Types I, III and IV
Repair systems • Proof-reading • Miss-match repair: Methyle directed missmatch repair • Excision repair:uvrA, B and C endonucleases
SOS repair:ssDNA stimulates rec A • Rec A downregulates lex A whic repress SOS genes (18 genes) • Error prone DNA repairing system
MUTATION and VARIATION • BActerial populations are not homgeneous • Mutations occurr randomly (Luria-Delbruck experiment – fluctuation test, 1943)
Mutants • Auxotrophs: Biochemically different from the parent (prototroph) • Resistant: Antimicrobial resistance • Spontaneous mutations approx. 1 in 1 million
Mutagenes • UV-light (TT-dimers formations Mutations, replication errors)