Document Detail


Complementation of the yeast deletion mutant DeltaNCE103 by members of the beta class of carbonic anhydrases is dependent on carbonic anhydrase activity rather than on antioxidant activity.
MedLine Citation:
PMID:  15096093     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In recent years, members of the beta class of CAs (carbonic anhydrases) have been shown to complement Delta NCE103, a yeast strain unable to grow under aerobic conditions. The activity required for complementation of Delta NCE103 by tobacco chloroplast CA was studied by site-directed mutagenesis. E196A (Glu196-->Ala), a mutated tobacco CA with low levels of CA activity, complemented Delta NCE103. To determine whether restoration of Delta NCE103 was due to residual levels of CA activity or whether it was related to previously proposed antioxidant activity of CAs [Götz, Gnann and Zimmermann (1999) Yeast 15, 855-864], additional complementation analysis was performed using human CAII, an alpha CA structurally unrelated to the beta class of CAs to which the tobacco protein belongs. Human CAII complemented Delta NCE103, strongly arguing that CA activity is responsible for the complementation of Delta NCE103. Consistent with this conclusion, recombinant NCE103 synthesized in Escherichia coli shows CA activity, and Delta NCE103 expressing the tobacco chloroplast CA exhibits the same sensitivity to H2O2 as the wild-type strain.
Authors:
Daniel Clark; Roger S Rowlett; John R Coleman; Daniel F Klessig
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  The Biochemical journal     Volume:  379     ISSN:  1470-8728     ISO Abbreviation:  Biochem. J.     Publication Date:  2004 May 
Date Detail:
Created Date:  2004-04-20     Completed Date:  2004-09-07     Revised Date:  2010-09-21    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  England    
Other Details:
Languages:  eng     Pagination:  609-15     Citation Subset:  IM    
Affiliation:
Boyce Thompson Institute for Plant Research, Tower Road, Ithaca, NY 14853, USA.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Antioxidants / metabolism*
Carbonic Anhydrase II / chemistry,  genetics,  metabolism
Carbonic Anhydrases / chemistry,  classification*,  genetics,  metabolism*
Chloroplasts / enzymology
Gene Deletion*
Genetic Complementation Test*
Humans
Hydrogen Peroxide / metabolism,  pharmacology
Hydrogen-Ion Concentration
Kinetics
Molecular Sequence Data
Mutagenesis, Site-Directed / genetics
Oxidants / metabolism,  pharmacology
Oxidative Stress / drug effects
Saccharomyces cerevisiae / enzymology*,  genetics,  metabolism
Tobacco / cytology,  enzymology,  genetics
Chemical
Reg. No./Substance:
0/Antioxidants; 0/Oxidants; 7722-84-1/Hydrogen Peroxide; EC 4.2.1.-/Carbonic Anhydrase II; EC 4.2.1.1/Carbonic Anhydrases
Comments/Corrections

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