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Localisation of the binding site for the initiating substrate on the RNA polymerase from Sulfolobus acidocaldarius
Author(s) -
Grachev Mikhail A.,
Mustaev Arkadij A.,
Zaychikov Evgeny F.,
Lindner Anton J.,
Hartmann Guido R.
Publication year - 1989
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(89)80746-x
Subject(s) - sulfolobus acidocaldarius , binding site , rna polymerase , substrate (aquarium) , chemistry , sulfolobus , polymerase , rna , biochemistry , biology , archaea , enzyme , gene , ecology
RNA polymerase from the archaebacterium Sulfolobus acidocaldarius was chemically modified with AMP o ‐formylphenyl ester followed by reduction with borohydride. The modified protein catalyzes the labeling of its own largest subunit when incubated with [α‐ 33 P]UTP in the presence of poly[d(A‐T)]. On cleaving of the labeled protein using cyanogen bromide, hydroxylamine or amino acid‐specific endoproteinases for a very brief period, the pattern and size of the radioactive fragments formed are best explained by attachment of the label between Gly 843 and Met 895 of the largest subunit. In this region there exists a highly conserved sequence which is also found in other archaebacterial, eukaryotic and prokaryotic RNA polymerases. This suggests that the binding site for the initiating substrate of RNA polymerases has been conserved during evolution.