Mapping the active site of yeast RNA polymerase B (II).
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.
Département de Biologie, Centre d'Etudes Nucléaires de Saclay, Gif-sur-Yvette, France.
M
M
Riva
C
C
Carles
A
A
Sentenac
M A
MA
Grachev
A A
AA
Mustaev
E F
EF
Zaychikov
D000595:Amino Acid Sequence; D001665:Binding Sites; D003488:Cyanogen Bromide; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases; D010449:Peptide Mapping; D012319:RNA Polymerase II / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid
D046911:Macromolecular Substances; D012319:RNA Polymerase II; D010447:Peptide Hydrolases; D003488:Cyanogen Bromide