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Originally published In Press as doi:10.1074/jbc.M108271200 on October 11, 2001

J. Biol. Chem., Vol. 276, Issue 51, 47922-47929, December 21, 2001
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Developmental Regulation of Heat Shock Protein 83 in Leishmania
3' PROCESSING AND mRNA STABILITY CONTROL TRANSCRIPT ABUNDANCE, AND TRANSLATION IS DIRECTED BY A DETERMINANT IN THE 3'-UNTRANSLATED REGION*

Alon ZilkaDagger , Srinivas GarlapatiDagger , Edit Dahan, Victoria Yaolsky, and Michal Shapira§

From the Department of Life Sciences, Ben Gurion University of the Negev, Beer-Sheva 84105, Israel

Developmental gene regulation in trypanosomatids proceeds exclusively by post-transcriptional mechanisms. Stability and abundance of heat shock protein (HSP)70 and HSP83 transcripts in Leishmania increase at mammalian-like temperatures, and their translation is enhanced. Here we report that the 3'-untranslated region (UTR) of HSP83 (886 nucleotides) confers the temperature-dependent pattern of regulation on a chloramphenicol acetyltransferase (CAT) reporter transcript. We also show that the majority of the 3'-UTR sequences are required for increasing mRNA stability during heat shock. Processing of the HSP70 and HSP83 primary transcripts to poly(A)+ mRNA was more efficient during heat shock; therefore, even when stability at 33 °C was reduced by deletions in the 3'-UTR, transcripts still accumulated to comparable and even higher levels. Translation of heat shock transcripts in Leishmania increases dramatically upon temperature elevation. Unlike in other eukaryotes in which the 5'-UTR confers preferential translation on heat shock transcripts, we show that translational control of HSP83 in Leishmania originates from its 3'-UTR. The 5'-UTR alone cannot induce translation during heat shock, but it has a minor contribution when combined with the HSP83 3'-UTR. We identified an element located between positions 201 and 472 of the 3'-UTR which is essential for increasing translation of the CAT-HSP83 reporter RNA at 33-37 °C. This region confers preferential translation during heat shock even in transcripts that were less stable. Thus, investigating the traditionally conserved heat shock response reveals that Leishmania parasites use unique pathways for translational control.


* This work was supported by German-Israel Binational Fund Grant I-350-062/94 and by Israel Science Foundation Grant 215/98.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger These authors contributed equally to this work.

§ To whom correspondence should be addressed: Dept. of Life Sciences, Ben Gurion University of the Negev, P. O. Box 653, Beer-Sheva 84105, Israel. Tel.: 972-8-647-2663; Fax: 972-8-647-2992; E-mail: shapiram@bgumail.bgu.ac.il.


Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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