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PMID: 2886907 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Isolation of human cDNAs for asparagine synthetase and expression in Jensen rat sarcoma cells.

Molecular and cellular biology ·Vol. 7 ·No. 7 ·1987-07-00 ·Pages 2435-43

Andrulis IL, Chen J, Ray PN

Abstract

Asparagine synthetase cDNAs containing the complete coding region were isolated from a human fibroblast cDNA library. DNA sequence analysis of the clones showed that the message contained one open reading frame encoding a protein of 64,400 Mr, 184 nucleotides of 5' untranslated region, and 120 nucleotides of 3' noncoding sequence. Plasmids containing the asparagine synthetase cDNAs were used in DNA-mediated transfer of genes into asparagine-requiring Jensen rat sarcoma cells. The cDNAs containing the entire protein-coding sequence expressed asparagine synthetase activity and were capable of conferring asparagine prototrophy on the Jensen rat sarcoma cells. However, cDNAs which lacked sequence for as few as 20 amino acids at the amino terminal could not rescue the cells from auxotrophy. The transferant cell lines contained multiple copies of the human asparagine synthetase cDNAs and produced human asparagine synthetase mRNA and asparagine synthetase protein. Several transferants with numerous copies of the cDNAs exhibited only basal levels of enzyme activity. Treatment of these transferant cell lines with 5-azacytidine greatly increased the expression of asparagine synthetase mRNA, protein, and activity.

MeSH Terms
Animals Aspartate-Ammonia Ligase/biosynthesis,genetics Base Sequence Cell Line DNA/genetics,isolation & purification Gene Expression Regulation Humans Ligases/genetics Rats Sarcoma, Experimental/enzymology,genetics Transcription, Genetic Transfection
Chemicals
DNA Ligases Aspartate-Ammonia Ligase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Andrulis I L
Chen J
Ray P N
References (29)
29 references, click to expand
  1. Suppression of murine leukemias by L-asparaginase. Incidence of sensitivity among leukemias of various types: comparative inhibitory activities of guinea pig serum L-asparaginase and Escherichia coli L-asparaginase.
    J Exp Med. 1967 Jan 1;125(1):17-31 PMID: 5334543
  2. The amino acid requirements of the Jensen sarcoma in vitro.
    Cancer Res. 1959 Jul;19(6, Part 1):591-5 PMID: 13671417
  3. Asparagine synthetase activity of mouse leukemias.
    Science. 1968 May 3;160(3827):533-5 PMID: 5689413
  4. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  5. Two types of ribosome in mouse-hamster hybrid cells.
    Nat New Biol. 1971 Mar 10;230(10):52-4 PMID: 5279808
  6. Detection of specific sequences among DNA fragments separated by gel electrophoresis.
    J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
  7. L-Asparaginase, vincristine, and prednisone for induction of first remission in acute lymphocytic leukemia.
    Cancer Res. 1977 Feb;37(2):535-40 PMID: 264412
  8. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I.
    J Mol Biol. 1977 Jun 15;113(1):237-51 PMID: 881736
  9. A role for asparaginyl-tRNA in the regulation of asparagine synthetase in a mammalian cell line.
    Proc Natl Acad Sci U S A. 1977 Jun;74(6):2367-9 PMID: 18735
  10. Asparaginyl-tRNA aminoacylation levels and asparagine synthetase expression in cultured Chinese hamster ovary cells.
    J Biol Chem. 1979 Nov 10;254(21):10629-33 PMID: 40971
  11. Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure.
    Nucleic Acids Res. 1979 Nov 24;7(6):1541-52 PMID: 503860
  12. Isolation and characterization of CHO cell mutants with altered asparagine synthetase.
    Somatic Cell Genet. 1979 Sep;5(5):625-39 PMID: 43600
  13. Chinese hamster ovary cells resistant to beta-aspartylhydroxamate contain increased levels of asparagine synthetase.
    J Biol Chem. 1980 May 25;255(10):4808-13 PMID: 6102991
  14. Elevated levels of asparagine synthetase activity in physiologically and genetically derepressed Chinese hamster ovary cells are due to increased rates of enzyme synthesis.
    J Biol Chem. 1981 Jul 25;256(14):7311-5 PMID: 6114097
  15. Purification of rat liver asparagine synthetase by affinity chromatography on reactive blue 2-agarose.
    Anal Biochem. 1981 Jun;114(1):163-6 PMID: 6116467
  16. DNA-mediated gene transfer of beta-aspartylhydroxamate resistance into Chinese hamster ovary cells.
    Proc Natl Acad Sci U S A. 1981 Sep;78(9):5724-8 PMID: 6117859
  17. Isolation and characterization of Chinese hamster ovary cell mutants resistant to the amino acid analog beta-aspartyl hydroxamate.
    Somatic Cell Genet. 1982 Jul;8(4):533-45 PMID: 6126933
  18. A cDNA cloning vector that permits expression of cDNA inserts in mammalian cells.
    Mol Cell Biol. 1983 Feb;3(2):280-9 PMID: 6300662
  19. Isolation and characterization of a full-length expressible cDNA for human hypoxanthine phosphoribosyl transferase.
    Proc Natl Acad Sci U S A. 1983 Jan;80(2):477-81 PMID: 6300847
  20. Chromosomal alterations associated with overproduction of asparagine synthetase in albizziin-resistant Chinese hamster ovary cells.
    Mol Cell Biol. 1983 Mar;3(3):391-8 PMID: 6133214
  21. Properties of asparagine synthetase in asparagine-independent variants of Jensen rat sarcoma cells induced by 5-azacytidine.
    Mol Cell Biol. 1983 Nov;3(11):1937-42 PMID: 6197624
  22. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs.
    Nucleic Acids Res. 1984 Jan 25;12(2):857-72 PMID: 6694911
  23. Isolation of functional cDNA clones for human thymidylate synthase.
    J Biol Chem. 1984 Dec 10;259(23):14361-4 PMID: 6094554
  24. Molecular cloning of a cDNA for Chinese hamster ovary asparagine synthetase.
    Gene. 1984 Oct;30(1-3):1-9 PMID: 6150880
  25. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
    Gene. 1985;33(1):103-19 PMID: 2985470
  26. Characterization of single step albizziin-resistant Chinese hamster ovary cell lines with elevated levels of asparagine synthetase activity.
    J Biol Chem. 1985 Jun 25;260(12):7523-7 PMID: 2860112
  27. Rapid and reliable dideoxy sequencing of double-stranded DNA.
    Gene. 1985;40(2-3):317-23 PMID: 3007296
  28. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  29. L-asparagine biosynthesis by nutritional variants of the Jensen sarcoma.
    Biochem Biophys Res Commun. 1967 Jan 23;26(2):228-33 PMID: 6067662
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.7.7.2435-2443.1987
Published
1987-07-00
Pages
2435-43
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC365375
Subset
IM
Databases
GENBANK
M16763
External Links
PubMed source DOI: 10.1128/mcb.7.7.2435-2443.1987 PMC Full Text View journal details
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