BIOL241 2013 Lecture 26: DNA repair

What type of mutations does the system fix? How does it recognize the lesion? What genes are involved? I) Proofreading DNA polymerases have proofreading activity ( !"#! exonuclease$% &n '% coli( DNA pol &&& has an error rate of )*"#% With proofreading( the error rate is reduced to )*"+% II) i!"atc# repair a$ DNA methylation is used to discriminate ,etween strands at a mismatch% -acteria contain dam methylase( which adds a methyl group to the adenine at the .A/0 se1uence% 2ollowing one round of replication( the newly synthesized strand remains temporarily unmethylated% At the mismatch site( he repair enzyme preferentially ,inds to the unmethylated strand% ,$ 'ndonuclease ma3es a nic3 either #! or ! to the mismatch on the unmethylated strand% c$ /he nic3ed DNA is unwound and degraded ,y an exonuclease% d$ DNA polymerase fills in the gap and ligase seals the nic3% e$ &n '% coli( 4ut H( 5( and 6 are involved in mismatch repair% 4ut 6 recognizes the initial mismatch% 4ut H and 5 proteins ma3e the nic3% f$ 6imilar mechanisms exist in yeast and in mammals% III) Po!t rep$ication repair%#o"o$ogou! reco"&ination repair (e%g% repairing /" dimers$ a$ 7ecA proteins directs a recom,inational exchange with the corresponding region on the undamaged parental strand (donor strand$% ,$ /he gap in the donor strand is filled ,y repair synthesis (ie( filled"in ,y DNA polymerase and sealed ,y ligase$% I') (O( repair (the last resort$ a$ activates the expressions of a,out 8* genes( which allow DNA replication to occur in the presence of these types of lesions% ,$ During 696 repair( DNA synthesis is error"prone( inserting random and possi,ly incorrect nucleotides% /hat is( 696 repair is mutagenic%

BIOL241 2013
') P#otoacti)ation repair reversal of :; damage in pro3aryotes% " <hotoreactivation enzyme (<7'$ a,sor,s a photo of light to cleave the ,onds ,etween thymine dimers% &n ,acteria( <7' is not essential% <7' has not ,een found in eu3aryotes% 'I) Ba!e e*ci!ion repair corrects damage to nitrogenous ,ases% a$ /he first step is that DNA glycosylases recognize chemically altered ,ases% Distinct DNA glycosylases can recognize different DNA damages% 2or instance( uracil"DNA glycosylase recognizes the presence of uracil in DNA% ,$ /he DNA glycosylases cut the glycosidic ,ond ,etween the ,ase and the sugar( creating an A< site% c$ /his sugar with a missing ,ase is recognized ,y A< endonuclease( which ma3es a cut in the phosphodiester ,ac3,one at the A< site% /his creates a distortion in DNA( which is then corrected% d$ Not clear if -'7 functions in eu3aryotes% 'II) Nuc$eotide e*ci!ion repair repairs ,ul3y lesions in DNA that alter or distort dou,le helix (e%g% /"dimers$% a$ &n '% coli( :vr genes (uvrA( uvr-( and uvr0$ recognize and clip out specific num,er of nucleotides around ,oth sides of the lesion% &n '% coli( a total of ) nucleotides( including the lesion( are removed% ,$ DNA polymerase and ligase then repair the gap% c$ &n human(disruption of the N'7 is associated with =eroderma pigmentosum (=<$( a rare recessive disorder that predisposes individuals to severe s3in a,normalities% When exposed to :; radiation (sun light$( =< individuals exhi,it reactions that range from frec3ling and s3in ulceration to the development of s3in cancer% 'III) Dou&$e+!trand &rea, repair repairs damages from ionizing radiation ,y reattaching two DNA strands a$ D6- can ,e repaired ,y homologous recom,inational repair% &n yeast( this re1uires 7AD#8 complex and occurs during late 6>.8 phase% ,$ D6- can also ,e repaired ,y non"homologous recom,inational repair( or end ?oining% /his system is activated in .)( prior to DNA replication% /his sytesm is error"prone%