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.
| Speed, Michael Clayton; Burkhart, Brett W; Picking, Jonathan W et al. (2018) An Archaeal Fluoride-Responsive Riboswitch Provides an Inducible Expression System for Hyperthermophiles. Appl Environ Microbiol 84: |
| 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 |
| Mattiroli, Francesca; Bhattacharyya, Sudipta; Dyer, Pamela N et al. (2017) Structure of histone-based chromatin in Archaea. Science 357:609-612 |
| Burkhart, Brett W; Cubonova, Lubomira; Heider, Margaret R et al. (2017) The GAN Exonuclease or the Flap Endonuclease Fen1 and RNase HII Are Necessary for Viability of Thermococcus kodakarensis. J Bacteriol 199: |
| Gehring, Alexandra M; Sanders, Travis J; Santangelo, Thomas J (2017) Markerless Gene Editing in the Hyperthermophilic Archaeon Thermococcus kodakarensis. Bio Protoc 7: |
| 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; Walker, Julie E; Santangelo, Thomas J (2016) Transcription Regulation in Archaea. J Bacteriol 198:1906-1917 |
| Gehring, Alexandra M; Santangelo, Thomas J (2015) Manipulating archaeal systems to permit analyses of transcription elongation-termination decisions in vitro. Methods Mol Biol 1276:263-79 |
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