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PMID: 6366788 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Androgen induction of ornithine decarboxylase mRNA in mouse kidney as studied by complementary DNA.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 81 ·No. 3 ·1984-02-00 ·Pages 731-5

Kontula KK, Torkkeli TK, Bardin CW, Jänne OA

Abstract

To investigate the mechanisms by which androgens regulate ornithine decarboxylase (OrnDCase; L-ornithine carboxy-lyase, EC 4.1.1.17) in mouse kidney, a cDNA clone encoding OrnDCase mRNA was prepared. Purification of OrnDCase mRNA from kidneys of androgen-treated mice was accomplished by immunoadsorption of renal polysomes to a protein A-Sepharose column and enrichment for poly(A)-containing RNA by oligo(dT)-cellulose. Double-stranded cDNA synthesized from this mRNA was inserted into the Pst I site of plasmid pBR322 by using oligo(dG . dC)-tailing and was propagated in Escherichia coli. Plasmids containing cDNA sequences coding for OrnDCase were identified by differential colony hybridization, by radioimmunological detection of OrnDCase-like antigens in bacterial cultures, and by cell-free translation of hybrid-selected mRNA followed by immunoprecipitation with monospecific OrnDCase antiserum. A restriction endonuclease fragment of the selected plasmid DNA (pODC54) was labeled by nick-translation and used to study changes in OrnDCase mRNA concentration. After a single dose of testosterone, renal OrnDCase mRNA concentration increased as soon as 6 hr and peaked 24 hr after steroid injection, as measured by RNA blot hybridization. Continuous androgen treatment for 4 days resulted in a 10- to 20-fold increase in OrnDCase mRNA concentration in normal animals, but no induction of this mRNA was detected in mice that have an inherent defect of the androgen receptor (testicular feminization). These results indicate that androgens regulate OrnD-Case synthesis in mouse kidney, at least in part, by increasing OrnDCase mRNA accumulation.

MeSH Terms
Animals Cloning, Molecular DNA/metabolism Drug Implants Enzyme Induction Escherichia coli/genetics Female Kidney/drug effects,metabolism Male Mice Mice, Inbred Strains Nucleic Acid Hybridization Ornithine Decarboxylase/genetics Plasmids RNA, Messenger/genetics Testosterone/pharmacology
Chemicals
Drug Implants RNA, Messenger Testosterone DNA Ornithine Decarboxylase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kontula K K
Torkkeli T K
Bardin C W
Jänne O A
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45 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.81.3.731
Published
1984-02-00
Pages
731-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC344909
Subset
IM
Grants
FIC NIH HHS · 3 FO5 TW3192-0151 · United States
NICHD NIH HHS · HD-13541 · United States
External Links
PubMed source DOI: 10.1073/pnas.81.3.731 PMC Full Text View journal details
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