Sequence-specific transcription termination by bacteriophage T7 RNA polymerase = 박테리오파아지 T7 RNA 중합효소의 염기서열 특이적 전사종결

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As well as canonical hairpin-forming terminator, phage RNA polymerases also recognize intrinsic, hairpin-independent termination signal. A conserved 7-base pair sequence (ATCTGTT in the non-template strand) and U-rich sequence downstream of it are required for termination. Mutations at four amino acid residues, K303, R307, E738 and D766, increase termination efficiency on at hairpin-independent termination signal. Guanidine hydrochloride increases the termination efficiency at hairpin-independent terminator but exerts no effect on hairpin terminator, suggesting that a conformational change promoted by the denaturant be involved in this termination process. We also found that termination could occur prior to the known termination site. At each of the termination sites, RNA polymerase complex manifests multiple states. When a normal elongation complex is trapped by the termination signal, it becomes a paused complex, which is still capable of resuming transcription. The paused complex becomes a termination complex that is destined to disintegrate. The paused complex could revert to normal elongation state. It can also proceed further via an alternative pathway, which is slow and requires high NTP concentration compared to normal elongation. Collapse of transcription bubble at a region corresponding to the conserved sequence was observed 4 bp before the termination site. From these results, we proposed a model for termination at hairpin-independent termination signal.
Advisors
Kang, Chang-Wonresearcher강창원researcher
Description
한국과학기술원 : 생물과학과,
Publisher
한국과학기술원
Issue Date
2001
Identifier
165780/325007 / 000965213
Language
eng
Description

학위논문(박사) - 한국과학기술원 : 생물과학과, 2001.2, [ v, 96 p. ]

Keywords

bacteriophage; transcription; 전사종결; 박테리오파아지; termination

URI
http://hdl.handle.net/10203/27502
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=165780&flag=dissertation
Appears in Collection
BS-Theses_Ph.D.(박사논문)
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