Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Disruption and overexpression of the Schizosaccharomyces pombe aph1 gene and the effects on intracellular diadenosine 5 ',5 '''-P-1,P-4-tetraphosphate (Ap(4)A), ATP and ADP concentrations
Autore:
Ingram, SW; Barnes, LD;
Indirizzi:
Univ Texas, Hlth Sci Ctr, Dept Biochem, San Antonio, TX 78229 USA Univ Texas San Antonio TX USA 78229 pt Biochem, San Antonio, TX 78229 USA
Titolo Testata:
BIOCHEMICAL JOURNAL
, volume: 350, anno: 2000,
parte:, 3
pagine: 663 - 669
SICI:
0264-6021(20000915)350:<663:DAOOTS>2.0.ZU;2-9
Fonte:
ISI
Lingua:
ENG
Soggetto:
SACCHAROMYCES-CEREVISIAE; ESCHERICHIA-COLI; BIS(5'-NUCLEOSIDYL) TETRAPHOSPHATES; AP4A PHOSPHORYLASE; TUMOR-SUPPRESSOR; FISSION YEAST; CELL-DIVISION; FHIT GENE; HYDROLASE; CLONING;
Keywords:
adenine nucleotides; diadenosine oligophosphates; fission yeast; gene manipulation;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
48
Recensione:
Indirizzi per estratti:
Indirizzo: Barnes, LD Univ Texas, Hlth Sci Ctr, Dept Biochem, Mail Code 7760,7703 Floyd Curl Dr,San Antonio, TX 78229 USA Univ Texas Mail Code 7760,7703 Floyd Curl Dr San Antonio TX USA 78229
Citazione:
S.W. Ingram e L.D. Barnes, "Disruption and overexpression of the Schizosaccharomyces pombe aph1 gene and the effects on intracellular diadenosine 5 ',5 '''-P-1,P-4-tetraphosphate (Ap(4)A), ATP and ADP concentrations", BIOCHEM J, 350, 2000, pp. 663-669

Abstract

Diadenosine oligophosphates are ubiquitous compounds that were discovered over 30 years ago. Diadenosine 5',5'''-P-1,P-4- tetraphosphate (Ap(4)A) is the most studied member of this family, and its function in yeast is unknown. To investigate possible functions, we changed the intracellular Ap(4)A concentration in Schizosaccharomyces pombe via disruption and overexpressionof the aph1 gene, which encodes an Ap(4)A hydrolase (Aph1). S. pombe Aph1 is 52% identical with a human tumour suppressor protein, Fhit, in a core region of 109 amino acids. Disruption of aph1 resulted in an 85% decrease in Ap(4)A hydrolase activity and a 290-fold increase in the intracellular Ap(4)A concentration. The disruption and subsequent increase in intracellular Ap(4)A concentration had no significant effect on the growth of S. pombe. Overexpression of the S. pombe aph1 gene, resulting in 17- and 84-fold increases in Ap(4)A hydrolase activity above wild-type levels, resulted in 60 and80% decreases respectively in the intracellular Ap(4)A concentration. Thisrepresents the first report of a decrease in the intracellular Ap(4)A concentration in response to overexpression of a degradative enzyme in any eukaryotic organism. We describe a new S. pombe expression plasmid, pPOX, whichwas used to achieve the largest increase in expression of aph1. Overexpression of aph1 at the highest level resulted in a 46% increase in generation time in comparison with the control strain. Neither overexpression nor disruption had any effect on the intracellular ATP or ADP concentrations. This is the first report of ADP and ATP concentrations in S. pombe. These data also indicate that Aph1 functions in vivo to degrade Ap(4)A, and that high-level overexpression of this enzyme reduces the growth rate.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 21/09/20 alle ore 11:36:36