?url_ver=Z39.88-2004&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Adc&rft.relation=http%3A%2F%2Fd-scholarship-dev.library.pitt.edu%2F21803%2F&rft.title=Assembly+and+Mechanism+of+Action+of+Sulfolobus+solfataricus+DNA+Replication+Complexes&rft.creator=Bauer%2C+Robert+J.&rft.description=DNA+replication+enzymes+are+essential+for+the+maintenance+and+propagation+of+genetic+information+which+precisely+governs+the+growth+and+development+of+our+cells.+Aberrant+DNA+replication+processes+have+been+implicated+in+a+wide+variety+of+human+diseases%2C+most+notably+cancer%2C+and+therefore%2C+mechanistic+understanding+of+DNA+replication+processes+is+paramount+for+the+development+of+human+therapeutic+agents.+The+study+of+the+eukaryotic+replication+system+however%2C+is+difficult%2C+as+the+system+contains+a+large+number+of+enzymes+and+regulatory+factors+making+assembly+of+these+systems+for+in+vitro+study+complicated.+Thus%2C+in+order+to+gain+insight+into+the+workings+of+the+eukaryotic+replication+system%2C+several+model+systems+are+used%2C+where+the+complexity+of+the+replication+pathways+is+not+as+great.%0D%0A%0D%0AThe+DNA+replication+system+from+the+thermophilic+archaeon+Sulfolobus+solfataricus+is+a+recently+identified+model+with+components+sharing+high+levels+of+sequence+homology+to+their+eukaryotic+counterparts.+This+system+is+ideal+for+gaining+insight+into+the+mechanistic+workings+of+DNA+replication+which+can+be+translated+to+the+eukaryotic+system.+A+key+advantage+to+the+study+of+thermophilic+enzymes+is+in+the+ability+to+utilize+reaction+temperatures+far+lower+than+the+physiological+conditions+for+the+organisms.+This+results+in+slower+kinetics+with+no+significant+change+in+overall+function%2C+allowing+an+easier+discernment+of+the+enzyme%E2%80%99s+mechanistic+details.+%0D%0A%0D%0AI+have+contributed+to+the+development+of+Sulfolobus+solfataricus+as+a+model+system+primarily+through+characterization+of+nucleotide+transferase+enzymes+including+DNA+polymerases+and+primases.+Firstly%2C+I+have+determined+that+the+DNA+polymerase%2C+SsoPolB3%2C+possesses+a+low+rate+of+synthesis+and+fidelity+more+similar+to+those+involved+in+lesion+bypass.+Secondly%2C+I+characterized+the+assembly+and+mechanism+of+action+SsoPolB1+replication+holoenzyme+which+replicates+in+a+distributive+fashion+similar+to+the+eukaryotic+Pold+holoenzyme%2C+and+maintains+stimulated+replication+rates+through+rapid+re-recruitment+of+the+polymerase+to+the+processivity+clamp.+Finally%2C+I+discovered+and+characterized+the+interactions+of+a+unique+primosome+complex+formed+between+the+bacterial+like+DnaG+primase+and+eukaryotic+like+MCM+helicase.+In+all%2C+my+thesis+provides+for+a+more+thorough+understanding+of+the+interactions%2C+kinetics%2C+and+dynamics+occurring+at+the+replication+fork.%0D%0A&rft.date=2014-09-16&rft.type=University+of+Pittsburgh+ETD&rft.type=PeerReviewed&rft.format=application%2Fpdf&rft.language=en&rft.identifier=http%3A%2F%2Fd-scholarship-dev.library.pitt.edu%2F21803%2F1%2FRobert_J_Bauer_Dissertation.pdf&rft.identifier=++Bauer%2C+Robert+J.++(2014)+Assembly+and+Mechanism+of+Action+of+Sulfolobus+solfataricus+DNA+Replication+Complexes.++Doctoral+Dissertation%2C+University+of+Pittsburgh.++++(Unpublished)++