Properties of rat and mouse beta-glucuronidase mRNA and cDNA, including evidence for sequence polymorphism and genetic regulation of mRNA levels

• Published in: Gene Vol. 36; no. 1-2; p. 15
• Main Authors: Watson, G, Felder, M, Rabinow, L, Moore, K, Labarca, C, Tietze, C, Vander Molen, G, Bracey, L, Brabant, M, Cai, J D
• Format: Journal Article
• Language: English
• Published: Netherlands 1985
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Cloning, Molecular; DNA - metabolism; Female; Genes; Glucuronidase - genetics; Kidney - enzymology; Mice; Mice, Inbred A; Mice, Inbred C57BL; Polymorphism, Genetic; Protein Biosynthesis; Rats; Rats, Inbred Strains; RNA, Messenger - genetics; Sebaceous Glands - enzymology; Species Specificity; Vagina - enzymology
• ISSN: 0378-1119
• DOI: 10.1016/0197-0186(85)90071-3

cDNA clones containing partial sequences for beta-glucuronidase (beta G) were constructed from rat preputial gland RNA and identified by their ability to selectively hybridize beta G mRNA. One such rat clone was used to isolate several cross-hybridizing clones from a mouse-cDNA library prepared from kidney RNA from androgen-treated animals. Together, the set of mouse clones spans about 2.0 kb of the 2.6-kb beta G mRNA. Using these cDNA clones as probes, a genomic polymorphism for DNA restriction fragment size was found that proved to be genetically linked to the beta G gene complex. A fragment of beta G cDNA was subcloned into a vector carrying an SP6 polymerase promoter to provide a template for the in vitro synthesis of single-stranded RNA complementary to beta G mRNA. This provided an extremely sensitive probe for the assay of beta G mRNA sequences. Using either nick-translated cDNA or transcribed RNA as a hybridization probe, we found that mouse beta G RNA levels are strongly induced by testosterone, and that induction by testosterone is pituitary-dependent. During the lag period preceding induction, during the induction period itself, and during deinduction following removal of testosterone, beta G mRNA levels paralleled rates of beta G synthesis previously measured by in vivo pulse-labelling experiments. Genetic variation in the extent of induction affected either the level of beta G mRNA or its efficiency of translation depending on the strain of mice tested.