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Stage-specific alternative splicing of the heat-shock transcription factor during the life-cycle of Schistosoma mansoni

Published online by Cambridge University Press:  05 October 2004

D. RAM ,
E. ZIV ,
F. LANTNER ,
V. LARDANS  and
I. SCHECHTER
D. RAM
Affiliation:
Department of Immunology, The Weizmann Institute of Science, Rehovot 76100, Israel
E. ZIV
Affiliation:
Department of Immunology, The Weizmann Institute of Science, Rehovot 76100, Israel
F. LANTNER
Affiliation:
Department of Immunology, The Weizmann Institute of Science, Rehovot 76100, Israel
V. LARDANS
Affiliation:
Department of Immunology, The Weizmann Institute of Science, Rehovot 76100, Israel
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Abstract

Stage-specific alternative splicing of the heat-shock transcription factor of Schistosoma mansoni (SmHSF) generates isoforms with structural diversity that may modulate the activity of SmHSF at different life-stages, and thus may regulate the expression of different genes at different developmental stages. RT-PCR, cloning and DNA-sequence analyses showed stage-specific alternative splicing inside the DNA-binding domain (DBD) involving introns I1 and I2, and beyond the DBD involving introns I4a and I7. Retention of introns I2 and I4a would inactivate SmHSF since they contain termination codons. Retention of intron I1 would add 11 amino acids inside the DBD and may change the DNA-binding specificity of SmHSF; intron I7 would add 13 amino acids to the effector region of HSF. Retention of introns was more pronounced in cercariae (larval stage living in water) than in adult worms (parasitic form in mammals). The isoforms were expressed in bacteria, but functional evaluation was not feasible, because only the isoform lacking introns was soluble while isoforms with introns were insoluble. However, stage-specific alternative splicing that changed HSF function in vivo was evidenced in intact cercariae. The cercarial SmHSF mRNA was enriched with introns I2 and I4a that contain termination codons. Therefore, translation of the SmHSF mRNA was impaired, and the SmHSF protein was undetectable. Consequently, the HSP70 gene could not be transcribed, and the HSP70 mRNA was missing. Alternative splicing was observed for short DNA segments (33–45 bp) bound by splice signals, located in the coding region. These are not bona fida exons since they are not flanked by introns. Yet, they are not regular introns since they are often found in mature mRNA. Alternative splicing of these DNA segments caused structural diversity that could modulate the function of the gene product.

Keywords

Schistosoma mansonialternative splicingheat-shock transcription factor isoformsstage-specific isoformsshort intronscontrol of stage-specific genes
Type
Research Article
Information
Parasitology , Volume 129 , Issue 5 , November 2004 , pp. 587 - 596
Copyright
© 2004 Cambridge University Press

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