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J. Biol. Chem., Vol. 265, Issue 27, 16498-16503, Sep, 1990
M Riva, C Carles, A Sentenac, MA Grachev, AA Mustaev and EF Zaychikov
Yeast RNA polymerase B (II) was incubated with a collection of 13 different
nucleotide derivatives and affinity labeled by allowing DNA- directed
phosphodiester bond formation. The 32P-labeled site was localized in the
C-terminal part of the B150 subunit by microsequencing a proteolytic
fragment, then further mapped by a combination of extensive or single-hit
chemical cleavage reactions and analysis of the labeled peptide patterns.
The affinity label was mapped to between Asn946 and Met999, within one of
the nine regions that are conserved between B150 and the bacterial beta
subunit. The results underscore the conservative evolution of the catalytic
center of eukaryotic and bacterial RNA polymerases.
Mapping the active site of yeast RNA polymerase B (II)
Departement de Biologie, Centre d'Etudes Nucleaires de Saclay, Gif-sur- Yvette, France.
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