Document Detail

A novel approach for expression cloning of small GTPases: identification, tissue distribution and chromosome mapping of the human homolog of rheb.
MedLine Citation:
PMID:  8543055     Owner:  NLM     Status:  MEDLINE    
We report a novel approach for identifying monomeric GTP-binding proteins that is based on probing cDNA expression libraries with [alpha-32P]GTP. In short, a nitrocellulose replica from a plated cDNA expression library is treated with 2% SDS to block the GTP-binding activity of various G proteins expressed by E. coli, thus allowing the direct identification of positive clones. Using this procedure we have cloned several small GTP-binding proteins from human keratinocytes including the human homolog of rheb, a novel member of the ras-related GTP-binding proteins. Human rheb cDNA shares 90% identity with the rat counterpart and it is highly upregulated in transformed human cells of various origin. Northern analysis showed that human rheb is ubiquitously expressed, with the highest levels observed in skeletal and cardiac muscle, and not in brain, as it is the case for rat rheb. The human RHEB gene was mapped to chromosome 10q11.
P S Gromov; P Madsen; N Tomerup; J E Celis
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  FEBS letters     Volume:  377     ISSN:  0014-5793     ISO Abbreviation:  FEBS Lett.     Publication Date:  1995 Dec 
Date Detail:
Created Date:  1996-02-13     Completed Date:  1996-02-13     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0155157     Medline TA:  FEBS Lett     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  221-6     Citation Subset:  IM    
Department of Medical Biochemistry, Aarhus University, Denmark.
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MeSH Terms
Amino Acid Sequence
Base Sequence
Cell Line, Transformed
Chromosome Mapping
Chromosomes, Human, Pair 10*
Cloning, Molecular
DNA, Complementary
GTP Phosphohydrolases / genetics*,  metabolism
GTP-Binding Proteins / chemistry,  genetics*,  metabolism
Gene Expression
Guanosine Triphosphate / metabolism
Molecular Sequence Data
Monomeric GTP-Binding Proteins*
Neuropeptides / chemistry,  genetics*,  metabolism
Phosphorus Radioisotopes
Reg. No./Substance:
0/DNA, Complementary; 0/Neuropeptides; 0/Phosphorus Radioisotopes; 0/RHEB protein, human; 0/Rheb protein, rat; 86-01-1/Guanosine Triphosphate; 9007-49-2/DNA; EC 3.6.1.-/GTP Phosphohydrolases; EC 3.6.1.-/GTP-Binding Proteins; EC GTP-Binding Proteins

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