Alanine-Scanning Mutagenesis of the Small-Subunit βA−βB Loop of Chloroplast Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase:  Substitution at Arg-71 Affects Thermal Stability and CO2/O2Specificity†

Biochemistry ◽  
2001 ◽  
Vol 40 (19) ◽  
pp. 5615-5621 ◽  
Author(s):  
Robert J. Spreitzer ◽  
Maria G. Esquivel ◽  
Yu-Chun Du ◽  
Patrick D. McLaughlin
Keyword(s):  
Thermal Stability ◽  
Small Subunit ◽  
Alanine Scanning Mutagenesis ◽  
Alanine Scanning ◽  
Bisphosphate Carboxylase

scholarly journals Analysis of the yeast transcription factor TFIIA: distinct functional regions and a polymerase II-specific role in basal and activated transcription.

1995 ◽  
Vol 15 (3) ◽  
pp. 1234-1243 ◽  
Author(s):  
J J Kang ◽  
D T Auble ◽  
J A Ranish ◽  
S Hahn
Keyword(s):  
Transcription Factor ◽  
Large Subunit ◽  
Small Subunit ◽  
Specific Factor ◽  
Alanine Scanning Mutagenesis ◽  
Alanine Scanning ◽  
Pol Ii ◽  
Conserved Regions

To probe the structure and function of the Saccharomyces cerevisiae general transcription factor TFIIA, we have systematically mutagenized the genes encoding both subunits and analyzed the effects of the mutations both in vivo and in vitro. We found that the central nonconserved region of the large subunit is not essential for function and likely acts as a spacer between the conserved N- and C-terminal regions. Deletion mutagenesis of the large subunit defined a region which is required for TATA binding protein (TBP) interaction. Alanine scanning mutagenesis defined a cluster of four basic residues which are likely required for interaction with DNA in the TBP-DNA complex. Much of the conserved regions of both subunits is required for subunit association, suggesting that these conserved regions fold into compact domains which extensively interact. In vitro transcription performed with extracts from yeast strains with mutations in either the large or the small TFIIA subunit demonstrated that TFIIA stimulates both basal and activated polymerase II (Pol II) transcription. The TFIIA-depleted extracts have normal Pol I and Pol III transcription activity, showing that TFIIA is a Pol II-specific factor. In vivo depletion of TFIIA activity reduced transcription from four different Pol II promoters. Finally, alanine scanning mutagenesis of TFIIA's small subunit has identified at least one mutation which is defective in transcription but which is not defective in subunit association or binding to TBP or TBP-DNA complexes.

scholarly journals Comprehensive Alanine-scanning Mutagenesis of Escherichia coli CsrA Defines Two Subdomains of Critical Functional Importance

2006 ◽  
Vol 281 (42) ◽  
pp. 31832-31842
Author(s):  
Jeffrey Mercante ◽  
Kazushi Suzuki ◽  
Xiaodong Cheng ◽  
Paul Babitzke ◽  
Tony Romeo
Keyword(s):  
Escherichia Coli ◽  
Functional Importance ◽  
Alanine Scanning Mutagenesis ◽  
Alanine Scanning

A short conserved sequence is involved in the light-inducibility of a gene encoding ribulose 1,5-bisphosphate carboxylase small subunit of pea

Nature ◽  
1985 ◽  
Vol 315 (6016) ◽  
pp. 200-204 ◽  
Author(s):  
Giorgio Morelli ◽  
Ferenc Nagy ◽  
Robert T. Fraley ◽  
Stephen G. Rogers ◽  
Nam-Hai Chua
Keyword(s):  
Small Subunit ◽  
Gene Encoding ◽  
Bisphosphate Carboxylase ◽  
Conserved Sequence ◽  
Light Inducibility
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