The Escherichia coli DnaK chaperone stimulates the α-complementation of β-galactosidase

Samuel Berhanu, Takuya Ueda, Jean Hervé Alix*

*この研究の対応する著者

研究成果: Article査読

抄録

pUC18 and pUC19 are well-known high copy-number plasmid vectors routinely used for DNA cloning purposes. We show here that, in Escherichia coli transformed by native pUC18, the α-complementation of β-galactosidase (i.e., mediated by the peptide LacZα18) is intrinsically weak and slow, but is greatly stimulated by the DnaK/DnaJ/GrpE chaperone system. In contrast, the α-complementation mediated by the peptide LacZα19 (in E. coli transformed by the native pUC19) is much more efficient and therefore does not require the assistance of the DnaK chaperone machinery. The marked difference between these two LacZα peptides is reproduced in a cell-free protein expression system coupled with α-complementation. We conclude that: (i) α-complementation of β-galactosidase is DnaK-mediated depending upon the LacZα peptide donor; (ii) DnaK, sensu stricto, is not necessary for α-complementation, but can enhance it to a great extent; (iii) this observation could be used to establish an easy and inexpensive method for screening small molecules libraries in search of DnaK inhibitors and also for deciphering the DnaK-mediated protein quality control mechanism.

本文言語English
ページ(範囲)669-688
ページ数20
ジャーナルJournal of Basic Microbiology
62
6
DOI
出版ステータスPublished - 2022 6月
外部発表はい

ASJC Scopus subject areas

  • 応用微生物学とバイオテクノロジー

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