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Author Wong, G. ♦ Kawajiri, K. ♦ Negishi, M.
Source United States Department of Energy Office of Scientific and Technical Information
Content type Text
Language English
Subject Keyword BASIC BIOLOGICAL SCIENCES ♦ LABELLED COMPOUNDS ♦ HYBRIDIZATION ♦ RECOMBINANT DNA ♦ DNA SEQUENCING ♦ TESTOSTERONE ♦ GENE REGULATION ♦ HYDROXYLASES ♦ MESSENGER-RNA ♦ MICE ♦ MICROSOMES ♦ OLIGONUCLEOTIDES ♦ PHOSPHORUS 32 ♦ SULFUR 35 ♦ ANDROGENS ♦ ANDROSTANES ♦ ANIMALS ♦ BETA DECAY RADIOISOTOPES ♦ BETA-MINUS DECAY RADIOISOTOPES ♦ CELL CONSTITUENTS ♦ DAYS LIVING RADIOISOTOPES ♦ DNA ♦ ENZYMES ♦ EVEN-ODD NUCLEI ♦ HORMONES ♦ HYDROXY COMPOUNDS ♦ ISOTOPES ♦ KETONES ♦ LIGHT NUCLEI ♦ MAMMALS ♦ NUCLEI ♦ NUCLEIC ACIDS ♦ ODD-ODD NUCLEI ♦ ORGANIC COMPOUNDS ♦ ORGANOIDS ♦ OXIDOREDUCTASES ♦ PHOSPHORUS ISOTOPES ♦ RADIOISOTOPES ♦ RNA ♦ RODENTS ♦ STEROID HORMONES ♦ STEROIDS ♦ STRUCTURAL CHEMICAL ANALYSIS ♦ SULFUR ISOTOPES ♦ VERTEBRATES 550201* -- Biochemistry-- Tracer Techniques
Abstract The cDNA clone p16..cap alpha..-1 for the male-specific isozyme (C-P-450/sub 16..cap alpha../)/sup 1/ of testosterone 16..cap alpha..-hydroxylase in livers of 129/J mice and two additional full-length cDNAs overlapping with p16..cap alpha..-1 (p16..cap alpha..-2 and p16..cap alpha..-16) were sequenced. p16..cap alpha..-2 contained a single open reading frame of 1512 nucleotides, consisting of 71 base pairs of the 5'-noncoding region and 63 base pairs of the 3'-noncoding region with an additional poly(A) tract. From this DNA sequence, C-P-450/sub 16..cap alpha../ was deduced to contain 504 amino acids with a calculated molecular mass of 56,948 daltons. p16..cap alpha..-1 showed a nucleotide sequence identical with that of p16..cap alpha..-2 but lacked nine amino acid residues from the N-terminus. Another cDNA clone, p16..cap alpha..-16, also exhibited the same coding sequence with the exception of a 142 base pair deletion spanning from nucleotide 853 to nucleotide 994 of p16..cap alpha..-2. The putative poly(A) addition signal AATAAA is present for all three clones, but there are polymorphisms in the start sites of polyadenylation. Amino acid alignment with P-450b, P-450 M1, and P-450c revealed that P-450/sub 16..cap alpha../ is 39.8%, 40.7%, and 31.5% homologous, respectively. Five highly homologous genes were selected from a genomic DNA library of BALB/cJ mice by plaque hybridization to the cDNA and characterized. The results indicated that p16..cap alpha..-1 recognizes at least two mRNAs in terms of their regulation by androgen; one is reversibly regulated and the other is neonatally imprinted in expression in adult livers. The two differentially regulated C-P-450/sub 16..cap alpha../ dependent testosterone 16..cap alpha..-hydroxylase activities in the liver microsomes from these mice were also detected and associated with the hybridizable mRNA levels. A hybridization of the specific oligonucleotide probe with the poly(A) + RNA concluded that p16..cap alpha..-1 and p16..cap alpha..-2 encode C-P-450/sub 16..cap alpha../, which is reversibly regulated.
Educational Use Research
Learning Resource Type Article
Publisher Date 1987-12-29
Publisher Place United States
Journal Biochemistry
Volume Number 26
Issue Number 26
Organization National Institute of Environmental Health Sciences, Research Triangle Park, NC (USA)


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