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This 10 page Class Notes was uploaded by AnnaClippinger on Monday September 28, 2015. The Class Notes belongs to BIOL-L211 2521 at Indiana University taught by Megan Dunn in Summer 2015. Since its upload, it has received 62 views. For similar materials see Molecular Biology in Biology at Indiana University.
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Date Created: 09/28/15
Tl l UT 3 Cl CN Step 24 2015 Lecture 9 Last Time Nucleic Acid Basic Methods Todav39 Hello mothers hello father M Dl cul lquot 151ml gq In line new Replication Initiation 39 Gl l39l39fLa Replication and the Mismatch Repair System Next Time Reading Assignment 39 quotCorrupt DNA might be good for you Chapter 10 pp 3149333 Chemical and Environmental Mutagens a Cellular Damage Mechanisms TLT um Dr False Double Stranded Brealcs Chondnq ha HQ E l if 0W 0 U N H tlTUt 1 Molecular biology in the news quotHello Mothers Hello Fatherquot The Economist Oct 2012 membrane bound or qonulu Qtno39ote m on M thi Celts AT P i how thtlr W n H bosomes Hewmemg f Wm H mm i UN in m r out o no mum M qontlu dole to SELF R ipll cala Cl pt 0 K am 0h E Ell that 239 3le 5 lm biotic wl 39 0m am not how 539 main thmm osumet t J qthm ALL mitochondria inheritedmaterhally dul Mg Hr it i l 1cm on Spam m Mi to ChUl39ldTl on them mm mm Md or Q39S39t39ruphquot m Mutations in mitochondrial DNA mtDNA can cause disease Wand climate 0 mm 1mm er lll ll MOleculor biology in the news quotHello Mothers Hello Father The Economist Oct 2012 ktpiqoooi mmle from MUM 5 Q WI WY mm mt no a WWW W l nu d to Y fmow a Nucleus mGWEd to a Emrmaj denueleaied Egg egg wl mutant M t DN H all Hi 1 0 Mitochondria withinMateo one Donnelea on Q Normalmit thtmd a Egoamnion to detain Wis 5 medal wn vow lrlll3ltl0l105ml1 l Ullt s House of Commons and Lords voted in favor of law allowing tthe parent babiesquot Molecular biology in the news First country to allow nVif39fg J 039 7 twin 3 mm l 1 h H leu IMUlSunng DNA Guided questions for reading quotHello Mothers Hello Fatherquot What is mtDNA How is it inherited What is meant by quotthree parent babies Describe mitochondrial transplantation Why was this technique developed w Dam m m know 013 M id th Implications OF Hquot O httpllwwwcnncoleDlSlDEfOBheaIthluh ivi B pemoIiibabies 4 Origins of replication 19 o a remain on J can i n Git t9 pitta n on M Mt than ONCE ifth Lg W LLR Known sequence L93 4m in tk WU i in humans Not all um no can Ncu Purim Ofthe ori used each is activated only ONCE per cell cycie V a No known consensns sequence I We mm at n dtiim when the m Hi I n is Una is no 5E1 Spn I We Willi focus on replication initiation i 39nly actnaiiy just E com In euikaryotes there are more proteins invi e Ci i8 cunning Wore Tmi om Dn new 5 Initiation ofDNA replication in E coli Driven by protein DNAandl proteinprotein interactions P 31 Yemen 0 the Adriatic 75 ripen or n E D nn A is activated m aw i DneArATP l 7 39 mn hm qW Iu Hi9 Hme e Oriqm in DNA strancis at 13 mer repeats separate DNAhelieaseHg i Mi i DNAheiicese 39 loader Enact ini aiion of DNA replication in coii d DnaB lhelicase at origin E 31 7 Emw 5 e DnaB recruits primase DNA palmerase Ill hnissnzwne i Sliding clamsz assemble on primers W is Delxxm m mi H aHs lanth on Mains mm 39 Llil ll l 395 initiation of DNA replication in E coii lagging Strands s dingdsmpl M 1311 quot 7 I L39 352 rquot L It in l p i It In 3 s Sliding clamps load on lagging strands DNA DUN mmse 17m l MHSWHTMHS 0n 1an mg warm h Two replication forks have been assembled quotInn TWO 0 replyNahum COW NW A llll A Growth and replication paradox in bacteria lFor rapidly growing cells it llakes as little as 20 m I 0n minutes to d ivid e But it takes at least 40 minutes to replicate the i I quot quot I r r 39 Chromosome 4 0 a 39 if a cell must have a chromosome how is it possible to dividefoster than replico on W a u quotan NIH PR rain Cali 0 an F0 m Multiple replication forks New rounds of replication begin before old round ends h r Time 1 PH om on i Dr L13 39 339 S39ClmE Olrigino i I f U i micamh Wtwn mate a m srur r Cilqu W mm m 53 Wilma 9 owllam mm 0 39 quotElmo Cl dug in W til I William UH new l DI1 can nq ClIromognme Ensures entire genome is replicated before cell division Eom bloc dol ill rmn tlrwli Mr K 39ChMWMDmt u mil 1W H quotmm will le um KL mg 4193 as l H DNA polymerase uses on exonucleose to proofread o resume DNAsyntheeie to removal of mismatched nuoleotitl39es mispairecl leetbase r pair 11 734 quot V I i 7 exunutilease aetive Ella j n m m our canquot ng 1 SWRSMQCH m the pa N my trust 7 removes improperly basespaired dNTPs m tint CE 1 nrtiot We of mama mun unitm But it isn t perfect and replication errors may remain 11 Replication errors and consequences op imantm enungt to Hit0N Itemtable 3 Si nt iris permanent Hquot patted to your off spring A nucleotide misincorporated during replication can become a mutation Mos t mnrlommutan39om will WSWHH l WES OF 1F it39mut Freed it 2 a wm toe pd SSEd an to 39 armme Second round replication i V first round d replication miss a 139 ineorporaliorr Fig 102 May alter protein sequence shape function A re mom mutation usually results in prote39 quot 39 39 mutation is extremely rare and increases or changes protein activity 12 39 r o quot i Jr39tr39 quotv39 I Wequot quot WWquot l l ll Replication errors Substitution mutatiens urnquot in DNA sequence caused be a single nucleotide base change insertion or deletion a an M an ltering of i Bum Clive mm a a bagel gran if if Mam on a time 39 replacement of a single Weleotide with another nucleotide tA 51 i PM Imidlntlo pwmldme Dr I H midi r1 Jfo lg1 we 0 Iquot Purim to PVTIMMME 395 T Q I Sequence of part nl39 a Sequence af 7 n unnal gene 7 mutated gene f iv z n EST 739 a Transition mutation in a1 ta EC in thls example an BEG to GEE In this example r quot 3 5 f rji 39v I H wt 539 39 39TrnT7 nr r 39 quot 5quot 13 Tzf wn Uri ni tne i FD ll39owmq 3 0 J 0m lg i n nam in on mum lm 5 added mii d r Addmnnnrremovalof one or mereanucleotldes bath purlrm e to s i ppa elDNA polymerase l GEE539 Replication errors Insertions or deletions in Often occur at repetitive sequences remember intern a nut 0r mm mm are aquot resut DNA DON meme Slipping It Sli pagalenditg togadd inn slippage leading tv ela m l i39 1 mm Minimum r r isllppagein 7 It Sii Bin ll l MO 9 i M at l l 339 WP 13 7h 17 MP W 3 New nmncl V WOW 5 WDMIW 0 l L new Stm nd 539 T mm in tm WWW 5 r 3 ecnnnnngetne 339 GGABABGTEQ ClLlU lN mom 4 f V W Slim 11 Fain nehllld lk Wan M template 5mm fc39 Tt tTt e39t iTG 5 m 339 Gunnnnneetnequot 339 eonnncetnc AA 14 39 39quot Hu Hi ton 5 SECJ39S E rt Mair or quot f Genetic disorders caused by an insertion of a greater than normal number of trinucleotidle repeatsin a speci c gene Hereditaryquot brain disorder 1 Results in progressive cognitive decline llwoluntary movements and loss of physical control quotGAGquot ood o n codes for glutamine ET f aquot 39 TrialM Eleni9 quotquot 31f39quotT quot wquotv r M39j quot39f39a CH 1 W lmdf le l 3W liabilm lduem suppage JW i Will name ins funnier oi the WOW 15 t coll Follows the replication rnachineriir llhmi dl qRN it Corrects mista kes 39 Marital autumnn 39 a thanqun nnwwur aounhelii iS DUllle L1 proteins warring f CDmplUl together i s a f Tt 1 Rm 9 n i s93 ll 5 iii 039 Changes conformation Requires ATP 15 Mismatch repair system in E coli cont cruitecl V V V milr EQlU nSurromdlmnm mmamn Mut it than 1 tr llqlgt strandled mm lullw N N Wrms 11m strand a lunwincls nicked strand i Pr cemnws DNH mm rm nice to the Mt or How f r quot quot in the gapsg39alled a 22 A l mismatch rapaired Fig 103 17 Mismatch repairsystemin EkEoli cont quotl 39WQ have to kmw Wth huatohdt w i t0 mmow 7 x iv nucleotide is replaced then 7 nucleotide will become permanently incarcerated How clues the mismatch repair system knew which of the two nucleotides to replace a H mgcrl m 1 m1 iEs hEl39w een e t m39plat Strum 1 tm mwm Wl l i h e i lEd Strand repaired Fig 10 3 18 E mli Dem methyla on at replication fork Dam m hqmgg line DNA is methylated at S HGATC B sequences by 7 I v f hum Prior to replicatinn m p is Witt th and m lthI Mandi fulw mtmlut l one Willth 9 1 AT C 5quot is a oi pram l H During DNA synthesis A 1 Newly synthesized strand is not 1 immediately methylated itMl qmwpi o m v m 9 m era M m f EHl 7 Fig 105 19 E coli Dam methylation at replication fark But how are the twe strands distinguished Mum T tLOQleE Moles the nun mumLatel mm39 Fig 10 5 What about mismatch repair in eukaryotes DO not have MUl tH Mun makes man i Do not use hemi methvlatien to mark new strands flr9quot Ulla f39l39 gl39 39 l l39S39 WSlldrng Clde W ldW T9 1le l Pall m 20 Hill quot
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