Localisation of the binding site for the initiating substrate on the RNA polymerase from Sulfolobus acidocaldarius

FEBS Letters

Grachev, Mikhail A.¹  Zaychikov, Evgeny F.¹  Lindner, Anton J.²  Hartmann, Guido R.²  Mustaev, Arkadij A.¹ 

[1] Limnological Institute, Siberian Division of the Academy of Sciences of the USSR, 664033 Irkutsk, USSR;Institut für Biochemie, Ludwig-Maximilians-Universität, D-8000 München 2, FRG

关键词: RNA polymerase;    Affinity labeling;    Active center;    Localization;    (Sulfolobus acidocaldarius);   

DOI  :  10.1016/0014-5793(89)80746-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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-³³P]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 Gly⁸⁴³ and Met⁸⁹⁵ 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.

【 授权许可】

Unknown