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J. Biol. Chem., Vol. 281, Issue 31, 21617-21628, August 4, 2006

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Two Frameshift Products Involved in the Transposition of Bacterial Insertion Sequence IS629^*

Chang-Chieh Chen and Shiau-Ting Hu¹

From the Institute of Microbiology and Immunology, School of Life Sciences and the Department of Microbiology, School of Medicine, National Yang-Ming University, No. 155, Li-Nong St., Sec. 2, Shih-Pai, Taipei 112, Taiwan

IS629 is 1,310 bp in length with a pair of 25-bp imperfect inverted repeats at its termini. Two partially overlapping open reading frames, orfA and orfB, are present in IS629, and two putative translational frameshift signals, TTTTG (T₄G) and AAAAT (A₄T), are located near the 3'-end of orfA. With the lacZ gene as the reporter, both T₄G and A₄T motifs are determined to be a –1 frameshift signal. Two peptides representing the two transframe products designated OrfAB' and OrfAB, are identified by a liquid chromatography-tandem mass spectrometric approach. Results of transposition assays show that OrfAB' is the transposase and that OrfAB aids in the transposition of IS629. Pulse-chase experiments and Escherichia coli two-hybrid assays demonstrate that OrfAB binds to and stabilizes OrfAB', thus increasing the transposition activity of IS629. This is the first transposable element in the IS3 family shown to have two functional frameshifted products involved in transposition and to use a transframe product to regulate transposition.

Received for publication, March 15, 2006 , and in revised form, May 8, 2006.

^* This work was supported by research grants (NSC90-2320-B-010-058 and NSC92-2320-B-010-063) from the R.O.C. National Science Council (to S.-T. H.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed. Tel.: 886-2-2826-7107; Fax: 886-2-2821-2880; E-mail: tingnahu{at}ym.edu.tw.

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