The reactivity of sulfhydryl groups of yeast DNA dependent RNA polymerase I

Paulina Bull, Ursula Wyneken, Pablo Valenzuela

Resultado de la investigación: Article

4 Citas (Scopus)

Resumen

The number of reactive cysteine residues of yeast RNA polymerase I was determined and their function was studied using parachloromercury benzoate(pCMB), dithiobisnitrobenzoate (DTNB) and N-ethyl-maleimide (NEM) as modifying agents. By treatment with DTNB about 45 sulfhydryl groups react in the presence of 8M urea. Under non-denaturing conditions only 20 sulfhydryl groups are reactive with pCMB and DTNB. Both reagents completely inactivate the enzyme and this effect can be reversed by reducing agents. The sedimentation coefficient and the subunit composition are not affected when the enzyme is inactivated. Two of the most reactive sulfhydryl groups are necessary for activity. The modification of these groups is partially protected by substrates and DNA, suggesting that they are involved either in catalysis or in the maintenance of the conformation of the active site. Experiments with 14C-NEM indicate that the most reactive groups are located in subunits of 185,000, 137,000 and 41,000 daltons.

Idioma originalEnglish
Páginas (desde-hasta)5149-5160
Número de páginas12
PublicaciónNucleic Acids Research
Volumen10
N.º17
DOI
EstadoPublished - 11 sep 1982

Huella dactilar

RNA Polymerase I
Benzoates
Reactivity
RNA
Yeast
DNA
Enzymes
Yeasts
Dependent
Reducing Agents
Reducing agents
Catalysis
Sedimentation
Urea
Cysteine
Conformations
Catalytic Domain
Maintenance
Substrates
Chemical analysis

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Citar esto

Bull, Paulina ; Wyneken, Ursula ; Valenzuela, Pablo. / The reactivity of sulfhydryl groups of yeast DNA dependent RNA polymerase I. En: Nucleic Acids Research. 1982 ; Vol. 10, N.º 17. pp. 5149-5160.
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The reactivity of sulfhydryl groups of yeast DNA dependent RNA polymerase I. / Bull, Paulina; Wyneken, Ursula; Valenzuela, Pablo.

En: Nucleic Acids Research, Vol. 10, N.º 17, 11.09.1982, p. 5149-5160.

Resultado de la investigación: Article

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T1 - The reactivity of sulfhydryl groups of yeast DNA dependent RNA polymerase I

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AU - Wyneken, Ursula

AU - Valenzuela, Pablo

PY - 1982/9/11

Y1 - 1982/9/11

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AB - The number of reactive cysteine residues of yeast RNA polymerase I was determined and their function was studied using parachloromercury benzoate(pCMB), dithiobisnitrobenzoate (DTNB) and N-ethyl-maleimide (NEM) as modifying agents. By treatment with DTNB about 45 sulfhydryl groups react in the presence of 8M urea. Under non-denaturing conditions only 20 sulfhydryl groups are reactive with pCMB and DTNB. Both reagents completely inactivate the enzyme and this effect can be reversed by reducing agents. The sedimentation coefficient and the subunit composition are not affected when the enzyme is inactivated. Two of the most reactive sulfhydryl groups are necessary for activity. The modification of these groups is partially protected by substrates and DNA, suggesting that they are involved either in catalysis or in the maintenance of the conformation of the active site. Experiments with 14C-NEM indicate that the most reactive groups are located in subunits of 185,000, 137,000 and 41,000 daltons.

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