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PMID: 3463967 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Nucleotide sequence of rat preputial gland beta-glucuronidase cDNA and in vitro insertion of its encoded polypeptide into microsomal membranes.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 19 ·1986-10-00 ·Pages 7292-6

Nishimura Y, Rosenfeld MG, Kreibich G, Gubler U, Sabatini DD, Adesnik M, Andy R

Abstract

We have selected the rat preputial gland beta-glucuronidase as a model protein to study the sorting of newly synthesized lysosomal hydrolases to the lysosome. The complete coding sequence of beta-glucuronidase messenger RNA was determined from the sequences of a group of overlapping cDNA clones isolated from preputial gland cDNA libraries. The beta-glucuronidase mRNA primary translation product contains 648 amino acids, including an amino-terminal signal sequence of 22 residues. The polypeptide has four potential sites for the addition of asparagine-linked core oligosaccharides. A 376-residue segment of beta-glucuronidase shows extensive homology (23% sequence identity) to a portion of Escherichia coli beta-galactosidase. This homology most likely reflects an evolutionary relationship between the bacterial and eukaryotic enzymes and the conservation of structural features necessary for the glycosidase activity of both proteins. Translation of mRNA synthesized in vitro by transcription of a cDNA containing the entire beta-glucuronidase coding region yielded a polypeptide that was immunoprecipitated with anti-beta-glucuronidase antiserum and had the same electrophoretic mobility as the primary translation product of natural beta-glucuronidase mRNA. In the presence of microsomal membranes, the in vitro-synthesized beta-glucuronidase underwent cotranslational incorporation into the microsomes, as indicated by removal of the signal sequence and the addition of several oligosaccharide chains. The beta-glucuronidase cDNA will provide a useful tool to study the mechanism of mannose phosphorylation and other aspects of the sorting of lysosomal enzymes to lysosomes.

MeSH Terms
Animals Cell-Free System Cloning, Molecular DNA/genetics Gene Expression Regulation Genitalia, Male/enzymology Glucuronidase/genetics,metabolism Glycosylation Male Membrane Proteins/genetics,metabolism Microsomes/enzymology Molecular Weight Protein Processing, Post-Translational Protein Sorting Signals/metabolism Rats
Chemicals
Membrane Proteins Protein Sorting Signals DNA Glucuronidase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Nishimura Y
Rosenfeld M G
Kreibich G
Gubler U
Sabatini D D
Adesnik M
Andy R
References (45)
45 references, click to expand
  1. Multiple lysosomal enzyme deficiency due to enzyme leakage?
    N Engl J Med. 1971 Jan 14;284(2):109-10 PMID: 4249768
  2. Purification of beta-glucuronidase from the preputial gland of the female rat.
    Biochem J. 1958 May;69(1):22-7 PMID: 13535577
  3. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA.
    Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110-4 PMID: 4559594
  4. Egasyn, a protein complexed with microsomal beta-glucuronidase.
    J Biol Chem. 1975 Feb 10;250(3):1146-8 PMID: 1112792
  5. Beta-glucuronidase of rat preputial gland. Crystallization, properties, carbohydrate composition, and subunits.
    J Biochem. 1975 Feb;77(2):427-38 PMID: 236293
  6. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis.
    Anal Biochem. 1976 Jan;70(1):75-85 PMID: 1259158
  7. Terminal labeling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase.
    Nucleic Acids Res. 1976 Apr;3(4):863-77 PMID: 775445
  8. Methionine 500, the site of covalent attachment of an active site-directed reagent of beta-galactosidase.
    J Biol Chem. 1978 Aug 10;253(15):5283-5 PMID: 97290
  9. A bacterial clone synthesizing proinsulin.
    Proc Natl Acad Sci U S A. 1978 Aug;75(8):3727-31 PMID: 358198
  10. Rapid purification of plasmid DNAs by hydroxyapatite chromatography.
    Eur J Biochem. 1978 Nov 2;91(1):303-10 PMID: 363426
  11. Early events in the biosynthesis of the lysosomal enzyme cathepsin D.
    J Biol Chem. 1979 Dec 10;254(23):11771-4 PMID: 500673
  12. A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
    Nucleic Acids Res. 1979 Nov 24;7(6):1513-23 PMID: 388356
  13. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease.
    Biochemistry. 1979 Nov 27;18(24):5294-9 PMID: 518835
  14. I-cell disease: deficiency of extracellular hydrolase phosphorylation.
    Biochem Biophys Res Commun. 1979 Dec 14;91(3):976-81 PMID: 526296
  15. Biosynthesis of lysosomal enzymes in fibroblasts. Phosphorylation of mannose residues.
    J Biol Chem. 1980 May 25;255(10):4946-50 PMID: 6989822
  16. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  17. Biosynthetic intermediates of beta-glucuronidase contain high mannose oligosaccharides with blocked phosphate residues.
    J Biol Chem. 1980 Jul 25;255(14):6633-9 PMID: 7391040
  18. Amino acid sequences containing cysteine or half-cystine residues in beta-glucuronidase.
    Aust J Biol Sci. 1980 Oct;33(5):513-20 PMID: 7213258
  19. Transcriptional control in the production of liver-specific mRNAs.
    Cell. 1981 Mar;23(3):731-9 PMID: 7226226
  20. Fibroblasts from patients with I-cell disease and pseudo-Hurler polydystrophy are deficient in uridine 5'-diphosphate-N-acetylglucosamine: glycoprotein N-acetylglucosaminylphosphotransferase activity.
    J Clin Invest. 1981 May;67(5):1574-9 PMID: 6262380
  21. Transport and processing of lysosomal enzymes by smooth muscle cells and endothelial cells.
    Exp Cell Res. 1981 May;133(1):23-30 PMID: 7238595
  22. The phosphorylation of beta-glucuronidase oligosaccharides in mouse P388D1 cells.
    J Biol Chem. 1981 Dec 25;256(24):13060-7 PMID: 7309750
  23. Biosynthesis of lysosomal hydrolases: their synthesis in bound polysomes and the role of co- and post-translational processing in determining their subcellular distribution.
    J Cell Biol. 1982 Apr;93(1):135-43 PMID: 7068751
  24. Cloning of a cDNA complementary to rat preputial gland beta-glucuronidase mRNA.
    Biochem Biophys Res Commun. 1982 Feb 26;104(4):1271-8 PMID: 7041909
  25. Demonstration of the heterozygous state for I-cell disease and pseudo-Hurler polydystrophy by assay of N-acetylglucosaminylphosphotransferase in white blood cells and fibroblasts.
    Am J Hum Genet. 1982 Sep;34(5):717-29 PMID: 6289658
  26. Inactivation of beta-Galactosidase by iodination of tyrosine-253.
    Biochemistry. 1982 Sep 28;21(20):5052-5 PMID: 6814483
  27. Amino acid sequence of porcine spleen cathepsin D light chain.
    J Biol Chem. 1983 May 25;258(10):6435-43 PMID: 6406481
  28. Homology of amino acid sequences of rat liver cathepsins B and H with that of papain.
    Proc Natl Acad Sci U S A. 1983 Jun;80(12):3666-70 PMID: 6574504
  29. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination.
    Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963-5 PMID: 6575390
  30. Carboxyl-terminal proteolytic processing during biosynthesis of the lysosomal enzymes beta-glucuronidase and cathepsin D.
    Biochemistry. 1983 Oct 25;22(22):5201-5 PMID: 6360205
  31. Detection of early changes in androgen-induced mouse renal beta-glucuronidase messenger ribonucleic acid using cloned complementary deoxyribonucleic acid.
    Biochemistry. 1983 Dec 20;22(26):6049-53 PMID: 6689271
  32. Identification of an essential carboxylate group at the active site of lacZ beta-galactosidase from Escherichia coli.
    Eur J Biochem. 1984 Feb 1;138(3):527-31 PMID: 6420154
  33. A simple and very efficient method for generating cDNA libraries.
    Gene. 1983 Nov;25(2-3):263-9 PMID: 6198242
  34. Genetic control of levels of murine kidney glucuronidase mRNA in response to androgen.
    Proc Natl Acad Sci U S A. 1983 Dec;80(24):7596-600 PMID: 6584873
  35. Amino acid sequence of porcine spleen cathepsin D.
    Proc Natl Acad Sci U S A. 1984 Jun;81(12):3703-7 PMID: 6587385
  36. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
    Nucleic Acids Res. 1984 Sep 25;12(18):7035-56 PMID: 6091052
  37. Molecular cloning of a cDNA for human alpha-L-fucosidase.
    Proc Natl Acad Sci U S A. 1985 Feb;82(4):1262-5 PMID: 2983333
  38. Rapid and sensitive protein similarity searches.
    Science. 1985 Mar 22;227(4693):1435-41 PMID: 2983426
  39. Identification of cDNA clones encoding a precursor of rat liver cathepsin B.
    Proc Natl Acad Sci U S A. 1985 Apr;82(8):2320-4 PMID: 2986112
  40. Isolation and expression in Escherichia coli of a cDNA clone encoding human beta-glucuronidase.
    Gene. 1985;34(1):105-10 PMID: 3924735
  41. Cloning and sequence analysis of cDNA for human cathepsin D.
    Proc Natl Acad Sci U S A. 1985 Aug;82(15):4910-4 PMID: 3927292
  42. Properties of rat and mouse beta-glucuronidase mRNA and cDNA, including evidence for sequence polymorphism and genetic regulation of mRNA levels.
    Gene. 1985;36(1-2):15-25 PMID: 3840761
  43. Human beta-hexosaminidase alpha chain: coding sequence and homology with the beta chain.
    Proc Natl Acad Sci U S A. 1985 Dec;82(23):7830-4 PMID: 2933746
  44. Nucleotide sequence of cDNA containing the complete coding sequence for human lysosomal glucocerebrosidase.
    J Biol Chem. 1986 Jan 5;261(1):50-3 PMID: 3001061
  45. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose.
    Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408-12 PMID: 4504350
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.83.19.7292
Published
1986-10-00
Pages
7292-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC386702
Subset
IM
Grants
NIA NIH HHS · AG 01461 · United States
NIGMS NIH HHS · GM 20277 · United States
Databases
GENBANK
M13962
External Links
PubMed source DOI: 10.1073/pnas.83.19.7292 PMC Full Text View journal details
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