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ID 17081
本文ファイル
著者
Ueno, Yoko
Kawamoto, Toru
Fujiwara, Tamaki
Komatsuzawa, Hitoshi
Oswald, Eric
キーワード
Cdt
lipoprotein
Actinobacillus actinomycetemcomitan
NDC
医学
抄録(英)
The cell cycle G2/M specific inhibitor, Cytolethal Distending Toxin (Cdt), from Actinobacillus actinomycetemcomitans is composed of CdtA, CdtB, and CdtC coded on the CdtA, CdtB and CdtC genes that are tandem on the chromosomal cdt locus. A. actinomycetemcomitans CdtA has the lipid binding consensus domain, the so-called “lipobox", at the N-terminal signal sequence. Using E.coli carrying pTK3022, we show the sixteenth residue, cysteine, of CdtA bound [3H]palmitate or [3H]glycerol. Further post translational processing of the signal peptide, CdtA, was inhibited using globomycin, an inhibitor of lipoprotein-specific signal peptidase II. Fractionation and immunoblot show the lipid modified CdtA is present in the outer membrane. Immuno-precipitation and the pull-down assay of the Cdt complex from E. coli carrying a plasmid containing CdtABC demonstrated the Cdt complex in the periplasm is composed of CdtA, CdtB, and CdtC, and that the Cdt complex in culture supernatant is a N-terminally truncated (36-43 amino acids) form of CdtA (CdtA'), CdtB, and CdtC. This suggests Cdt is present as a complex both in the periplasm and the supernatant where CdtA undergoes post-translation processing to CdtA' in the process of biogenesis and secretion of Cdt holotoxin into the culture supernatant. Site-directed mutagenesis of the sixteenth cysteine residue to glycine in CdtA altered localization of CdtA in the cell and reduced the amount of Cdt activity in the culture supernatant. This suggests Cdt forms a complex inside the periplasm for lipid modification where post-translational processing of CdtA plays an important role for the efficient production of Cdt holotoxin into the culture supernatant.
掲載誌名
Infection and Immunity
74巻
6号
開始ページ
3480
終了ページ
3487
出版年月日
2006-06
出版者
American Society for Microbiology
ISSN
0019-9567
NCID
出版者DOI
PubMedID
言語
英語
NII資源タイプ
学術雑誌論文
広大資料タイプ
学術雑誌論文
DCMIタイプ
text
フォーマット
application/pdf
著者版フラグ
author
権利情報
Copyright (c) 2006 American Society for Microbiology.
関連情報URL(IsVersionOf)
http://dx.doi.org/10.1128/IAI.00739-05
部局名
医歯薬学総合研究科