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Nucleic Acids Research, 1988, Vol. 16, No. 20 9567-9585
© 1988


Articles

Structure and expression of the hsp 70 gene family of Leishmania major

Mary Gwo-Shu Lee, Brent Lee Atkinson, Suzanne Holmes Giannini1 and Lex H.T. Van der Ploeg*

Department of Genetics and Development, Columbia University College of Physicians and Surgeons 701 West 168th Street, New York, NY 10032 1Department of Medicine and Center for Vaccine Development, University of Maryland School of Medicine Baltimore, MD 21201, USA

*To whom correspondence should be addressed

Received July 11, 1988. Accepted September 2, 1988.

The parasitic protozoan Leishmania major differentiates in vitro. from the insect-adapted promastigote to the mammallan infective amastigote, In response to a temperature shift from 25°C to 37°C. We studied the genes encoding 70 kilodalton heat shock proteins (hsp 70 genes) in Leishmania substocks, which vary in their capability to differentiate. In total, four hsp 70 genes are arranged in tandem with intergenic regions of about 380 bp. These hsp 70 genes are 89%ed conserved at the aminoacid level when compared to the T.brucei hsp 70 genes. The expression of these four hsp 70 genes is increased, in vitro and in vivo, in response to a temperature shift from 25°C to 37°C. The parasite thus indeed responds to the transfer between hosts like it responds to a heat shock. In contrast, the high rate of transcription of a fifth identical hsp 70 gene, located at a separate locus, is unaffected by temperature shifts. The hsp 70 mRNAs have mini-exons trans-spliced onto their 5' ends and share unusually long (1000 nt) 3' untranslated extensions containing repetitive sequences. It is unclear whether or not the Intergenic regions of the L.major hsp 70 genes function in transcription initiation and/or whether transcription results in the generation of polycistronic pre-mRNAs. Since each of the hsp 70 genes that we identified is expressed normally in an L.major substock that lost the capability to differentiate in response to an in vitro temperature shift, the inability to differentiate does not result from a general defect in the temperature-dependent control of transcription.


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