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Cell cycle regulation of HMG-CoA red...
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Held, Paul Glenn.
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Cell cycle regulation of HMG-CoA reductase.
紀錄類型:
書目-電子資源 : Monograph/item
正題名/作者:
Cell cycle regulation of HMG-CoA reductase./
作者:
Held, Paul Glenn.
面頁冊數:
230 p.
附註:
Source: Dissertation Abstracts International, Volume: 51-04, Section: B, page: 1650.
Contained By:
Dissertation Abstracts International51-04B.
標題:
Biology, Molecular. -
電子資源:
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9024660
Cell cycle regulation of HMG-CoA reductase.
Held, Paul Glenn.
Cell cycle regulation of HMG-CoA reductase.
- 230 p.
Source: Dissertation Abstracts International, Volume: 51-04, Section: B, page: 1650.
Thesis (Ph.D.)--Albany Medical College of Union University, 1990.
Initial experiments involving the measurement of HMG-CoA Reductase (HMGR) activity revealed that enzyme activity was dependent on cell seeding. Peak HMGR activity proceeds DNA synthesis and drops to low levels during S phase. Experimentation showed that cell attachment was time dependent and when monolaver cells were allowed to attach for short periods of time (5 min); those cells that attached behaved in a synchronous manner, with peak DNA synthesis occurring 16-18 hours following attachment. Comparison with other established synchronization methods revealed that the method isolates an early G1 population of cells as well as produce a similar degree of synchrony.Subjects--Topical Terms:
1017719
Biology, Molecular.
Cell cycle regulation of HMG-CoA reductase.
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Source: Dissertation Abstracts International, Volume: 51-04, Section: B, page: 1650.
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Thesis (Ph.D.)--Albany Medical College of Union University, 1990.
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Initial experiments involving the measurement of HMG-CoA Reductase (HMGR) activity revealed that enzyme activity was dependent on cell seeding. Peak HMGR activity proceeds DNA synthesis and drops to low levels during S phase. Experimentation showed that cell attachment was time dependent and when monolaver cells were allowed to attach for short periods of time (5 min); those cells that attached behaved in a synchronous manner, with peak DNA synthesis occurring 16-18 hours following attachment. Comparison with other established synchronization methods revealed that the method isolates an early G1 population of cells as well as produce a similar degree of synchrony.
520
$a
Using this synchronization procedure, we have demonstrated the requirement for mevalonate or a metabolite for DNA synthesis. DNA synthesis was restored by the addition of exogenous mevalonate when CHO cells were seeded in the presence of inhibitors of HMGR. Mevalonate was shown to be required 10-12 hours prior to the peak in DNA synthesis. Transcription of the HMGR gene is required in G1 as the effectiveness of 25-hydroxycholesterol, as an inhibitor of DNA synthesis, is lost when added to synchronized cultures after 6 hours from seeding. Cell cycle dependent expression of HMGR was investigated using attachment synchrony. RNA isolated from AMR-2 and CHO cells show increased levels 1-2 hours after seeding. This peak in mRNA is presumably the result of increased transcription of the gene.
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Chimeric fusions with the bacterial CAT gene indicate that regulatory elements are contained in nucleic acid sequences located in the promoter region and within the 3
$\
sp\prime
$-
untranslated region of HMGR mRNA. Synchronized CHO cells transfected with a CAT reporter plasmid containing the HMGR promoter show a pattern of CAT activity that parallels that of HMGR activity. CHO cells transfected with the SV2CATRED plasmid show a much lower activity that parallels the parent plasmid suggesting the presence of negative regulatory elements.
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