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Two different aspects of cell metabo...
~
Sabina, Jeffrey.
Two different aspects of cell metabolism: (i) Recovery from growth inhibition in vivo, and (ii) the biochemical consequences of a split gene.
紀錄類型:
書目-電子資源 : Monograph/item
正題名/作者:
Two different aspects of cell metabolism: (i) Recovery from growth inhibition in vivo, and (ii) the biochemical consequences of a split gene.
作者:
Sabina, Jeffrey.
面頁冊數:
170 p.
附註:
Director: Dieter G. Soll.
附註:
Source: Dissertation Abstracts International, Volume: 66-11, Section: B, page: 5966.
Contained By:
Dissertation Abstracts International66-11B.
標題:
Chemistry, Biochemistry.
電子資源:
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3194706
ISBN:
9780542394768
Two different aspects of cell metabolism: (i) Recovery from growth inhibition in vivo, and (ii) the biochemical consequences of a split gene.
Sabina, Jeffrey.
Two different aspects of cell metabolism: (i) Recovery from growth inhibition in vivo, and (ii) the biochemical consequences of a split gene.
- 170 p.
Director: Dieter G. Soll.
Thesis (Ph.D.)--Yale University, 2005.
Secondly, a directed biochemical study of the archaeal tRNA-guanine transglycosylase (TGT) from Pyrococcus furiosus determined that its recognition of guanine at position 15 in tRNA is localized to the catalytic core, a feature shared with its bacterial counterpart. The archaeal enzyme is 61% larger than the bacterial TGT, due to an additional 230 amino acids at its C-terminus. Truncated forms of P. furiosus TGT retain their specificity for guanine exchange at position 15. Deletion of the PUA domain caused a 4-fold drop in the observed catalytic rate (2.8 x 10-3 sec -1) and resulted in a 75-fold increased KM for tRNAAsp (1.2 x 10-5 M) compared to wild-type TGT. Mutations in tRNAAsp which alter interactions with the PUA domain or prevent guanine exchange, efficiently competed with wild-type tRNAAsp for binding to full-length and truncated TGTs. While the C-terminal domains do not appear to play a role in modification site selection, their relevance for in vivo function remains to be discovered.
ISBN: 9780542394768Subjects--Topical Terms:
226900
Chemistry, Biochemistry.
Two different aspects of cell metabolism: (i) Recovery from growth inhibition in vivo, and (ii) the biochemical consequences of a split gene.
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Secondly, a directed biochemical study of the archaeal tRNA-guanine transglycosylase (TGT) from Pyrococcus furiosus determined that its recognition of guanine at position 15 in tRNA is localized to the catalytic core, a feature shared with its bacterial counterpart. The archaeal enzyme is 61% larger than the bacterial TGT, due to an additional 230 amino acids at its C-terminus. Truncated forms of P. furiosus TGT retain their specificity for guanine exchange at position 15. Deletion of the PUA domain caused a 4-fold drop in the observed catalytic rate (2.8 x 10-3 sec -1) and resulted in a 75-fold increased KM for tRNAAsp (1.2 x 10-5 M) compared to wild-type TGT. Mutations in tRNAAsp which alter interactions with the PUA domain or prevent guanine exchange, efficiently competed with wild-type tRNAAsp for binding to full-length and truncated TGTs. While the C-terminal domains do not appear to play a role in modification site selection, their relevance for in vivo function remains to be discovered.
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The first approach was to monitor Escherichia coli's responses on a genome-wide scale to four inhibitors that interfere with translation using DNA microarrays and real-time PCR. 4-Azaleucine, a competitive inhibitor of leucyl-tRNA synthetase, triggered the heat shock response. Administration of mupirocin, an inhibitor of isoleucyl-tRNA synthetase, resulted in changes reminiscent of the stringent response. Treatment with kasugamycin and puromycin, targeting 70S ribosome assembly as well as its peptidyl-transferase activity, caused accumulation of mRNAs from ribosomal protein operons. Exposure of a relA- strain to mupirocin resulted in accumulation of ribosomal operon transcripts. However, the relA - strain's response to the other inhibitors was quite similar to that of the wild-type strain. Overall, the data provide insight into the metabolic reorganization that occurs in E. coli in response to stress.
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Translation involves the coordination of many cellular processes; from amino acid biosynthesis, to tRNA processing, to ribosome assembly. These pathways are finely controlled so that the components involved are present at adequate levels to meet the cell's demand for protein synthesis at any given moment. This large conglomeration of subsystems can be studied in aggregate, but a specific step can also be examined, with both approaches yielding compelling observations.
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