Characteristic function of the conserved sequence on the sequence-specific termination by T7 RNA polymerase = 파아지 T7 RNA 중합효소의 염기서열 특이적 전사종결에서 보존서열의 기능에 대한 연구

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A sequence-specific termination signal of E. coli operon rrnB t1 recognized by bacteriophage RNA polymerases consists of conserved upstream-module (ATCTGTT in the non-template strand) and U-rich downstream-module. We previously elucidated the formation of paused conformation just prior to the point of RNA release. In this study, we observed sequential characteristic changes of ternary complex from more previous to the termination site. Elongation complexes of T7 RNA polymerase were stopped at desired positions by stepwise walking utilizing complex immobilization via biotin-streptavidin conjugation. The significant 9-nt upstream contact observed by photocross-link between nascent RNA and RNA polymerase in the elongation mode remarkably diminished from the third nucleotide position of the upstream-module. The release rate of the RNA in ternary complex was abruptly increased just before the conserved sequence and further increased near the point of RNA release if DNA templates contained intact upstream-module. Templates mutated termination signal showed that upstream-module appeared to contribute characteristic changes to cross-linking pattern and complex stability. At a few bp before the RNA release site, the slow intermediate complexes requiring high NTP concentration was observed. The collapse of transcription bubble was shown prior to termination. The normal transcription bubble shrank in the complex possessed intact upstream-module termination signal. The sequential changes in elongating complex before termination suggested that termination is prepared from 15bp before the RNA release site and accompanied by the various conformational changes.
Advisors
Kang, Chang-Wonresearcher강창원researcher
Description
한국과학기술원 : 생명과학과,
Publisher
한국과학기술원
Issue Date
2004
Identifier
237519/325007  / 000995203
Language
eng
Description

학위논문(박사) - 한국과학기술원 : 생명과학과, 2004.2, [ vi, 111 p. ]

Keywords

CONSERVED SEQUENCE; SEQUENCE-SPECIFIC TERMINATION; T7 RNA POLYMERASE; TRANSCRIPTION; 전사; 보존서열; 염기서열 특이적 전사종결; 파아지 리보핵산 중합효소

URI
http://hdl.handle.net/10203/27555
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=237519&flag=dissertation
Appears in Collection
BS-Theses_Ph.D.(박사논문)
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