title: Post-transcriptional repair of a split heat shock protein 90 gene by mRNA trans-splicing.
abstract: Heat shock protein 90 participates in diverse biological processes ranging from protein folding, cell cycle, signal transduction and development to evolution in all eukaryotes. It is also critically involved in regulating growth of protozoa such as Dictyostelium discoideum, Leishmania donovani, Plasmodium falciparum, Trypanosoma cruzi, and Trypanosoma evansi. Selective inhibition of Hsp90 has also been explored as an intervention strategy against important human diseases such as cancer, malaria, or trypanosomiasis. Giardia lamblia, a simple protozoan parasite of humans and animals, is an important cause of diarrheal disease with significant morbidity and some mortality in tropical countries. Here we show that the G. lamblia cytosolic hsp90 (glhsp90) is split in two similar sized fragments located 777 kb apart on the same scaffold. Intrigued by this unique arrangement, which appears to be specific for the Giardiinae, we have investigated the biosynthesis of GlHsp90. We used genome sequencing to confirm the split nature of the giardial hsp90. However, a specific antibody raised against the peptide detected a product with a mass of about 80 kDa, suggesting a post-transcriptional rescue of the genomic defect. We show evidence for the joining of the two independent Hsp90 transcripts in-trans to one long mature mRNA presumably by RNA splicing. The splicing junction carries hallmarks of classical cis-spliced introns, suggesting that the regular cis-splicing machinery may be sufficient for repair of the open reading frame. A complementary 26-nt sequence in the "intron" regions adjacent to the splice sites may assist in positioning the two pre-mRNAs for processing. This is the first example of post-transcriptional rescue of a split gene by trans-splicing.
authors.affiliation: Department of Biochemistry, Indian Institute of Science, Bangalore 560012, India.
authors.forename: Rishi Kumar
authors.initials: RK
authors.lastname: Nageshan
authors.forename: Nainita
authors.initials: N
authors.lastname: Roy
authors.forename: Adrian B
authors.initials: AB
authors.lastname: Hehl
authors.forename: Utpal
authors.initials: U
authors.lastname: Tatu
mesh_terms: D000906:Antibodies / Q000494:pharmacology; D000918:Antibody Specificity; D016678:Genome; D016829:Giardia lamblia / Q000235:genetics*; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics* / Q000276:immunology; D015800:Protozoan Proteins / Q000235:genetics* / Q000276:immunology; D012322:RNA Precursors / Q000235:genetics; D012323:RNA Processing, Post-Transcriptional / Q000502:physiology*; D012326:RNA Splicing / Q000502:physiology*; D016053:RNA, Protozoan / Q000235:genetics
chemical_list: D000906:Antibodies; D018841:HSP90 Heat-Shock Proteins; D015800:Protozoan Proteins; D012322:RNA Precursors; D016053:RNA, Protozoan