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Transcription of alpha- and beta-tubulin genes in vitro in isolated Chlamydomonas reinhardi nuclei.
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MedLine Citation:
PMID:  6699084     Owner:  NLM     Status:  MEDLINE    
Removal of the flagella of Chlamydomonas results in increases in both flagellar protein synthesis and tubulin messenger RNA accumulation. These observations led us to examine whether flagellar protein gene sequences are transcribed differentially in nuclei isolated before and after deflagellation. A nuclear isolation protocol was developed using the cell wall-less strain of Chlamydomonas, CW 15, after cell lysis with 0.5% Nonidet P-40. Transcriptional activity of isolated nuclei was determined by incorporating [32P]UTP into TCA-precipitable and phenol-extractable RNA, and by hybridizing newly transcribed RNA to complementary DNA clones containing alpha- and beta-tubulin sequences. Nuclei from deflagellated cells are more active in transcribing sequences that hybridize with alpha- and beta-tubulin complementary DNA probes than are nuclei from nondeflagellated cells. In addition, while total [32P]UTP incorporation is inhibited 45% by alpha-amanitin concentrations of 1.0 micrograms/ml, tubulin RNA synthesis in this system is completely inhibited by this concentration of alpha-amanitin. This demonstration of differential transcription in nuclei before and after cell deflagellation provides the means to study in vitro the mechanisms that signal and regulate flagellar protein gene activity.
L R Keller; J A Schloss; C D Silflow; J L Rosenbaum
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The Journal of cell biology     Volume:  98     ISSN:  0021-9525     ISO Abbreviation:  J. Cell Biol.     Publication Date:  1984 Mar 
Date Detail:
Created Date:  1984-04-24     Completed Date:  1984-04-24     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0375356     Medline TA:  J Cell Biol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1138-43     Citation Subset:  IM    
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MeSH Terms
Cell Nucleus / physiology
Cell-Free System
Chlamydomonas / genetics*
Flagella / physiology*
Gene Expression Regulation
Transcription, Genetic
Tubulin / genetics*
Grant Support
Reg. No./Substance:

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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Journal Information
Journal ID (nlm-ta): J Cell Biol
ISSN: 0021-9525
ISSN: 1540-8140
Publisher: The Rockefeller University Press
Article Information
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Print publication date: Day: 1 Month: 3 Year: 1984
Volume: 98 Issue: 3
First Page: 1138 Last Page: 1143
ID: 2113143
Publisher Id: 84135981
PubMed Id: 6699084

Transcription of alpha- and beta-tubulin genes in vitro in isolated Chlamydomonas reinhardi nuclei

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