Open AccessYeast seryl‐tRNA synthetase expressed in Escherichia coli recognizes bacterial serine‐specific tRNAs in vivo
Author(s) -
WEYGANDDURAŠEVIĆ Ivana,
BAN Nenad,
JAHN Dieter,
SÖLL Dieter
Publication year - 1993
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1993.tb17990.x
Subject(s) - escherichia coli , complementation , yeast , biochemistry , saccharomyces cerevisiae , transfer rna , serine , biology , mutant , in vivo , microbiology and biotechnology , chemistry , gene , enzyme , rna , genetics
The Saccharomyces cerevisiae serS gene which encodes seryl‐tRNA synthetase (SerRS) was expressed in Escherichia coli from the promoter and the ribosome binding sequences contained in its own 5′‐flanking region. The low level of yeast SerRS in the prokaryotic host was sufficient to permit in vivo complementation of two temperature‐sensitive E. coli serS mutants at the nonpermissive temperature. Thus, yeast SerRS can aminoacylate E. coli tRNA Ser species in vivo . Yeast SerRS, isolated from an overexpressing E. coli strain by a rapid two‐step purification on FPLC, aminoacylated E. coli tRNA with serine much more poorly (relative k cat /K m = 2 × 10 −4 ) than its homologous tRNAs. DL‐Serine hydroxamate, an inhibitor of E. coli SerRS, inhibits yeast SerRS in vivo and in vitro with an inhibition constant ( K 1 ) of 2.7 mM, a value 90‐fold higher than that for E. coli SerRS.