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Volume 272, Number 13, Issue of March 28, 1997 pp. 8466-8473
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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Post-translational Processing and Renal Expression of Mouse Indian Hedgehog

(Received for publication, November 4, 1996)

Rudolph P. Valentini , William T. Brookhiser ^§ , John Park ^¶ , Tianxin Yang , Josephine Briggs , Gregory Dressler ^§** and Lawrence B. Holzman

From the Departments of  Pediatrics,  Internal Medicine, ^¶ Pediatric Urology, and ^** Pathology, and the ^§ Howard Hughes Medical Institute, University of Michigan Medical School, Ann Arbor, Michigan 48109-0676

The full-length mouse Indian hedgehog (Ihh) cDNA was cloned from an embryonic 17.5-day kidney library and was used to study the post-translational processing of the peptide and temporal and spatial expression of the transcript. Sequence analysis predicted two putative translation initiation sites. Ihh translation was initiated at both initiation sites when expressed in an in vitro transcription/translation system. Expression of an Ihh mutant demonstrated that the internal translation initiation site was sufficient to produce the mature forms of Ihh. Ihh post-translational processing proceeded in a fashion similar to Sonic and Drosophila hedgehog; the unprocessed form underwent signal peptide cleavage as well as internal proteolytic processing to form a 19-kDa amino-terminal peptide and a 26-kDa carboxyl-terminal peptide. This processing required His³¹³ present in a conserved serine protease motif. Ihh transcript was detected by in situ RNA hybridization as early as 10 days postcoitum (dpc) in developing gut, as early as 14.5 dpc in the cartilage primordium, and in the developing urogenital sinus. In semiquantitative reverse transcription-polymerase chain reaction experiments, Indian hedgehog transcript was first detected in the mouse metanephros at 14.5 dpc; transcript abundance increased with gestational age, becoming maximal in adulthood. In adult kidney, Ihh transcript was detected only in the proximal convoluted tubule and proximal straight tubule.

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