Molecular cloning and characterization of two rat renal kallikrein genes.
Kallikreins compose a multigene family coding for a subgroup of serine proteases, which are involved in the processing of bioactive peptides. Two rat kallikrein-related genes, RSKG-7 (rat submandibular gland kallikrein gene 7) and RSKG-3, have been cloned and their sequences analyzed. RSKG-7 is approximately 4200 bases in length and consists of five exons and four introns. The 5' end region contains the variant CATAT box and TTTAAA box; the 3' end region contains the polyadenylation signal AATAAA. This gene encodes a putative 28,935-dalton preproenzyme of 261 amino acids (aa). The active enzyme consists of 237 aa and is preceded by a deduced signal peptide of 18 aa and a profragment of 6 aa. RSKG-3 is highly homologous to RSKG-7 in terms of its sequence and structure; it encodes a 28,730-dalton prepropeptide consisting of a signal peptide of 18 aa, a profragment of 6 aa, and an active peptide of 235 aa. Sequence comparisons of RSKG-7, RSKG-3, and other kallikrein-related enzymes reveal the key amino acid residues needed for both serine protease activity (His/Asp/Ser) and kallikrein-like cleavage specificity at basic amino acids. Northern blot analyses using specific oligonucleotide probes demonstrate that, among the 12 tissues studied, RSKG-7 and RSKG-3 are expressed in the rat kidney and submandibular gland. Castration of male rats results in a decrease in submandibular gland RSKG-7 mRNA, which can be restored to the normal level by treatment with thyroxine or testosterone. On the other hand, neither castration nor hormonal manipulation affects RSKG-7 mRNA levels in the kidney.
Chen
Y P
YP
Department of Biochemistry, Medical University of South Carolina, Charleston 29425.
Chao
J
J
Chao
L
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D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D004262:DNA Restriction Enzymes; D005091:Exons; D005260:Female; D007438:Introns; D007610:Kallikreins / Q000235:genetics*; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009919:Orchiectomy; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013363:Submandibular Gland / Q000187:drug effects / Q000378:metabolism; D013739:Testosterone / Q000494:pharmacology; D013974:Thyroxine / Q000494:pharmacology; D014018:Tissue Distribution; D014158:Transcription, Genetic
D003062:Codon; D012333:RNA, Messenger; D013739:Testosterone; D004262:DNA Restriction Enzymes; D007610:Kallikreins; D013974:Thyroxine