Timelyandaccurateregulationofgeneexpressionisrequiredforpropergrowth,development, andresponsetoenvironmentalstimuli.Acompleteunderstandingofthemechanisms employedtoregulategeneexpressionisnecessarytocombattheaberrantregulationthat underliesmanyhumandevelopmentalconditionsandailments,includingcancer.Thelong-term goalsofthisproposalaretodeterminethemechanicsofRNAsynthesisbyDNA-dependent multi-subunitRNApolymerases(RNAPs),theregulationimposedonRNAPbyconserved proteinfactors,selecttemplatesequencesandchromatin-barriers,andtospecifically characterizetheavailablemechanismstohaltRNAsynthesisandterminatetranscription.The proposedexperimentstakeadvantageofanestablishedinvivoandinvitroarchaeal transcriptionsystemfromthemarinehyperthermophilicorganismThermococcuskodakarensis. Archaeaoffertheadvantagesoffarlesscomplexitybuthomologyinmanyfeaturesofhuman molecularbiology,specificallyconservationofRNAPstructureandfunctionandhistone-based chromatin.Wewillcharacterizefactor-dependentandintrinsictranscriptiontermination, polarity,theconsequencesofDNAdamageontranscriptionelongationandtermination,andthe roleofglobaltranscriptionregulationinarchaealphysiology.Theresultsobtainedwill determinethestructuresandsequencesthatsupporttranscriptionelongationcomplexstability, describethemechanicsofterminationandallowcomparisonsofterminationmechanismsin eachdomainoflife,andprovideinsighttocontrolandultimatelymanipulategeneexpressionto counteravarietyofdiseasesandailmentsaffectinghumanhealth.
Acompleteunderstandingofthemechanismsemployedtoregulategeneexpressionis necessarytocombattheaberrantregulationthatunderliesmanyhumandevelopmental conditionsandailments,includingcancer.Thelong-termgoalsofthisproposalareto determinethemechanicsofRNAsynthesisbyDNA-dependentmulti-subunitRNApolymerases (RNAPs)andtospecificallycharacterizetheavailablemechanismstohaltRNAsynthesisand terminatetranscription.Theresultsobtainedwilldeterminethestructuresandsequencesthat supporttranscriptionelongationcomplexstability,describethemechanismsofterminationand allowcomparisonsofterminationmechanismsineachdomain.
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| Walker, Julie E; Luyties, Olivia; Santangelo, Thomas J (2017) Factor-dependent archaeal transcription termination. Proc Natl Acad Sci U S A 114:E6767-E6773 |
| Gehring, Alexandra M; Santangelo, Thomas J (2017) Archaeal RNA polymerase arrests transcription at DNA lesions. Transcription 8:288-296 |
| Wang, Peng; Yu, Zhuoteng; Santangelo, Thomas J et al. (2017) BosR Is A Novel Fur Family Member Responsive to Copper and Regulating Copper Homeostasis in Borrelia burgdorferi. J Bacteriol 199: |
| Gehring, Alexandra M; Astling, David P; Matsumi, Rie et al. (2017) Genome Replication in Thermococcus kodakarensis Independent of Cdc6 and an Origin of Replication. Front Microbiol 8:2084 |
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