Preparation and characterization of a sex-dependent rat urinary protein☆

Roy, Arun K.; Neuhaus, Otto W.; Harmison, Charles R.

doi:10.1016/0304-4165(66)90477-6

Cited by 110 publications

(39 citation statements)

References 12 publications

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“…It has been reported that the only amino sugar in the glycosylated form of a2u-globulin is N-acetylglucosamine (11,12).…”

Section: And Discussionmentioning

confidence: 99%

“…5, black dot). A single base change at this site in another expressed a2u-globulin gene would explain the existence of both glycosylated and nonglycosylated forms of this protein (11,12). An analysis of this region of the amino acid chain by the predictive scheme of Chou and Fasman (44) indicates that this asparagine residue is contained in a sequence (residues 34-37) that exhibits a strong potential to exist in 8-turn conformation (45).…”

Section: And Discussionmentioning

confidence: 99%

“…This regulation in vivo has been shown to occur via modulation of the rate of synthesis of hepatic a2u-globulin mRNA (5, 10). The existence of both glycosylated and nonglycosylated forms of a2,-globulin has been reported (11,12), as well as induction of the glycosylated form by glucocorticoids (13,14). a2u-Globulin is first synthesized as the 21,000-dalton precursor pro-a2u-globulin, which is then processed to the mature form of the protein (15).…”

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confidence: 99%

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Cloning and sequence of several alpha 2u-globulin cDNAs.

Unterman¹,

Lynch²,

Nakhasi³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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We describe a simple cloning procedure for a%.-globulin that requires neither enrichment of mRNA for cloning nor purification of a specific probe for screening recombinant colonies. Total adult male liver poly(A)+RNA was used as template for cloning, and the subsequent recombinant colonies were screened by comparing hybridization to radioactive cDNA probes prepared from hepatic male and female mRNA, respectively. Almost all of the selected "male-specific" clones were later shown to contain a2,-globulin sequences. This cloned au-globulin cDNA has been shown to specifically hybridize to male rat liver RNA, which, when isolated and translated in vitro, codes for a 21,000-dalton protein (pro-a2u-globulin) immunologically identical to auglobulin. When translation occurs in the presence of pancreatic microsomes this in vitro synthesized pro-a2u-globulin is processed to the 19,000-dalton mature form of a2u-globulin. The nucleotide sequence of the a2u-globulin cDNA has been determined, thus elucidating the complete amino acid sequence of a2.-globulin and most of the hydrophobic "leader" sequence of pro-a2u-globulin.The amino acid sequence deduced from the cDNA is in agreement with the partial sequence that we previously determined by sequential Edman degradation ofthe purified-protein. a2u-Globulin cDNA clones contain within the.3'-untranslated region one or both of the two putative polyadenylylation/transcription termination sites (A-A-T-A-A-A and A-A-T-T-A-A-A). Either of these can be used, generating a2u-globulin mRNA species of two lengths. A codon usage analysis of the cDNA showed that, although all six leucine codons are used for the 14 leucine residues in mature auglobulin, the seven leucines in the partial leader sequence reported are all encoded by the same codon, CTG. The primary amino acid sequence contains a unique Asn-Gly-Ser sequence, likely to be in (-turn conformation, as the probable site of glycosylation for this glycoprotein.ac2u-Globulin is synthesized in the liver of normal adult male rats, secreted into the bloodstream, and excreted in the urine (1), and represents ':'1% of total hepatic protein synthesis (2). Female liver, male kidney, and several hepatomas do not synthesize a2u-globulin and have no detectable a2,,-globulin mRNA sequences (2-4). The biosynthesis of a2u-globulin is also under complex multihormonal control: androgens, glucocorticoids, thyroid hormone, insulin, and growth hormone stimulate and estrogens repress its synthesis (5-9). This regulation in vivo has been shown to occur via modulation of the rate of synthesis of hepatic a2u-globulin mRNA (5, 10). The existence of both glycosylated and nonglycosylated forms of a2,-globulin has been reported (11,12), as well as induction of the glycosylated form by glucocorticoids (13,14). a2u-Globulin is first synthesized as the 21,000-dalton precursor pro-a2u-globulin, which is then processed to the mature form of the protein (15).As part of our ongoing research into the regulation of a2u-globulin synthesis, we have cloned the cDNA ...

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“…It has been reported that the only amino sugar in the glycosylated form of a2u-globulin is N-acetylglucosamine (11,12).…”

Section: And Discussionmentioning

confidence: 99%

Section: And Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Cloning and sequence of several alpha 2u-globulin cDNAs.

Unterman¹,

Lynch²,

Nakhasi³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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“…The system involves androgen-dependent synthesis of a low molecular weight (23,000) protein called a2u globulin (10,11). We have also identified the mRNA for a2u globulin in rat liver and have established a strong correlation between the androgen-dependent increase in a2u synthesis and the hepatic concentration of the mRNA for this protein (9,12,13 Cycloheximide, cordycepin, and a-amanitin were administered in 0.14 M NaCl.…”

mentioning

confidence: 98%

Superinduction of alpha 2u globulin by actinomycin D: evidence for drug-mediated increase in alpha 2u mRNA.

Chatterjee

Hopkins

Dutchak

et al. 1979

Proc. Natl. Acad. Sci. U.S.A.

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Actinomycin D, an inhibitor of DNA-dependent RNA synthesis, increased the hepatic concentration of a2u globulin, an androgen-inducible protein in the rat. Spayed female rats with a marginally induced state of a2u synthesis showed an approximately 5-fold increase in hepatic a2u globulin within 3-6 hr after treatment with actinomycin D. Initial treatment of these animals with 5a-dihydrotestosterone, followed by actinomycin D, resulted within 2-3 hr in a more than 2-fold increase in hepatic a2u globulin compared to animals treated with the androgen alone. In spite of inhibition of hepatic synthesis of poly(A)containing RNA to less than 25% of control, superinduction with actinomycin D resulted in a parallel increase in the translatable mRNA for a2u globulin. These results showing increase in both a2u globulin and its translatable mRNA after superinduction with actinomycin D support the concept of post-transcriptional repression of a2u synthesis. Actinomycin D, an inhibitor of DNA-dependent RNA synthesis, increases the level of several inducible proteins, a phenomenon termed "superinduction" (1, 2). Because actinomycin D inhibits mRNA synthesis, superinduction by this drug indicates an apparent paradox, the resolution of which may provide important clues concerning the mechanism of gene regulation in the eukaryotic cell. In fact, the post-transcriptional model for the steroidal regulation of gene activity, proposed by Tomkins and coworkers, is based primarily on their interpretation of the superinduction of glucocorticoid-mediated synthesis of tyrosine aminotransferase (TAT) by actinomycin D in hepatoma cells and in rat liver (3). A similar model of translational repression was also suggested by McAuslan (4). The above investigators have proposed that the mRNAs for the inducible proteins are continually being synthesized within the permissive phase of the cell cycle. However, the simultaneous presence of rapidly turning over post-transcriptional repressors prevents the translation of these mRNAs and may lead to their degradation. It is also proposed that, in the presence of inducing steroids, the repressor is inactivated and, because of its short half-life, is preferentially eliminated by actinomycin D, both of which phenomena would favor an accumulation of the inducible mRNA leading to increased synthesis of the inducible protein (3). Due to technical limitations, experimental verification of the McAuslan-Tomkins model has been difficult, and various other interpretations for the phenomenon of superinduction by actinomycin D have been proposed (5-7).In our laboratory we have developed a model system for the study of hormone action in rat liver (8, 9). The system involves androgen-dependent synthesis of a low molecular weight (23,000) protein called a2u globulin (10, 11). We have also identified the mRNA for a2u globulin in rat liver and have established a strong correlation between the androgen-dependent increase in a2u synthesis and the hepatic concentration of the mRNA for this protein (9,12,13 Cyclohexim...

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“…This protein is synthesized in the liver, secreted into the blood stream, concentrated in the kidneys, and excreted in the urine. Its synthesis is under the synergistic control of androgens and glucocorticoids (3)(4)(5)(6)(7)(8)(9)(10)(11).…”

Section: At(11-1)-1754 Coo-1754-12mentioning

confidence: 99%

Effects of whole-body $gamma$-irradiation on the biosynthesis of certain serum proteins. Progress report, September 1, 1972--December 31, 1973

Neuhaus¹

1973

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Preparation and characterization of a sex-dependent rat urinary protein☆

Cited by 110 publications

References 12 publications

Cloning and sequence of several alpha 2u-globulin cDNAs.

Cloning and sequence of several alpha 2u-globulin cDNAs.

Superinduction of alpha 2u globulin by actinomycin D: evidence for drug-mediated increase in alpha 2u mRNA.

Effects of whole-body $gamma$-irradiation on the biosynthesis of certain serum proteins. Progress report, September 1, 1972--December 31, 1973

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