Molecular cloning of the human gene, CCG2, that complements the BHK-derived temperature-sensitive cell cycle mutant tsBN63: Identity of CCG2 with the human X chromosomal SCAR/RPS4X gene

Minoru Watanabe, Nobuaki Furuno, Mark Goebl, Mitiko Go, Kumi Miyauchi, Takeshi Sekiguchi, Claudio Basilico, Takeharu Nishimito

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

A temperature-sensitive mutant tsBN63 cell line was isolated by the fluorodeoxyuridine method from the BHK21/13 cell line after mutagenesis with nitrosoguanidine. When cultures of tsBN63 cells growing asynchronously at 33.5°C were shifted to 39.5°C, a nonpermissive temperature, the ability for protein synthesis was rapidly reduced and cell proliferation stopped mainly at G1 phase, and partly at G2 phase. Synchronized cultures of tsBN63 cells did not commence DNA synthesis when shifted up in G1 phase. The human gene complementing the tsBN63 mutation was cloned by DNA-mediated gene transfer and its cDNA of 1.1 kb conferring ts+ phenotype on tsBN63 cells was isolated from the cDNA library of Raj (mer+) cells with a frequency of 10-3. On the basis of the determined nucleotide sequence, the isolated human gene turned out to be the X chromosomal RPS4X encoding the ribosomal protein S4. The size of the CCG2 gene was estimated to be about 12kb by complementation analysis of the tsBN63 mutation with cloned genomic DNA.

Original languageEnglish
Pages (from-to)35-43
Number of pages9
JournalJournal of Cell Science
Volume100
Issue number1
StatePublished - Sep 1991

Fingerprint

Molecular Cloning
Cell Cycle
Temperature
G1 Phase
Genes
DNA
Cell Culture Techniques
Floxuridine
Nitrosoguanidines
Cell Line
Mutation
G2 Phase
Gene Library
Mutagenesis
Complementary DNA
Cell Proliferation
Phenotype
Proteins

Keywords

  • Cell cycle mutation
  • RPS4
  • tsBN63
  • X chromosome

ASJC Scopus subject areas

  • Cell Biology

Cite this

Molecular cloning of the human gene, CCG2, that complements the BHK-derived temperature-sensitive cell cycle mutant tsBN63 : Identity of CCG2 with the human X chromosomal SCAR/RPS4X gene. / Watanabe, Minoru; Furuno, Nobuaki; Goebl, Mark; Go, Mitiko; Miyauchi, Kumi; Sekiguchi, Takeshi; Basilico, Claudio; Nishimito, Takeharu.

In: Journal of Cell Science, Vol. 100, No. 1, 09.1991, p. 35-43.

Research output: Contribution to journalArticle

Watanabe, Minoru ; Furuno, Nobuaki ; Goebl, Mark ; Go, Mitiko ; Miyauchi, Kumi ; Sekiguchi, Takeshi ; Basilico, Claudio ; Nishimito, Takeharu. / Molecular cloning of the human gene, CCG2, that complements the BHK-derived temperature-sensitive cell cycle mutant tsBN63 : Identity of CCG2 with the human X chromosomal SCAR/RPS4X gene. In: Journal of Cell Science. 1991 ; Vol. 100, No. 1. pp. 35-43.
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abstract = "A temperature-sensitive mutant tsBN63 cell line was isolated by the fluorodeoxyuridine method from the BHK21/13 cell line after mutagenesis with nitrosoguanidine. When cultures of tsBN63 cells growing asynchronously at 33.5°C were shifted to 39.5°C, a nonpermissive temperature, the ability for protein synthesis was rapidly reduced and cell proliferation stopped mainly at G1 phase, and partly at G2 phase. Synchronized cultures of tsBN63 cells did not commence DNA synthesis when shifted up in G1 phase. The human gene complementing the tsBN63 mutation was cloned by DNA-mediated gene transfer and its cDNA of 1.1 kb conferring ts+ phenotype on tsBN63 cells was isolated from the cDNA library of Raj (mer+) cells with a frequency of 10-3. On the basis of the determined nucleotide sequence, the isolated human gene turned out to be the X chromosomal RPS4X encoding the ribosomal protein S4. The size of the CCG2 gene was estimated to be about 12kb by complementation analysis of the tsBN63 mutation with cloned genomic DNA.",
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