A gene correcting the defect in the CHO mutant Ade-H, deficient in a branch point enzyme (adenylosuccinate synthetase) of de novo purine biosynthesis, is located on the long arm of chromosome 1

Li-Wen Lai, Iris M. Hart, David Patterson

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Somatic hybrids between human cells and the Chinese hamster ovary (CHO) K1 mutant, Ade-H cells, were selected for purine prototrophy by growth in adenine-free medium. The Ade-H mutant is defective in the enzyme adenylosuccinate (AMPS) synthetase (ADSS; EC 6.3.4.4), which carries out the first of a two-step sequence in the biosynthesis of AMP from IMP, and therefore requires exogenous adenine for growth. The presence of the long arm of human chromosome 1 in the hybrids is 100% concordant for the ability to growth in adenine-free medium and restoration of the enzyme activity. Hybrid segregants that lose the ability to grow in adenine-free medium lose all or a portion of chromosome 1 and enzyme activity. Southern blot hybridization with a chromosome 1-specific probe, BCMI, confirms the existence of human chromosome 1 in these hybrids. Analysis of a human/CHO translocation chromosome that arose in one of the hybrids suggests that the gene correcting the defect lies in the region 1 cen-1q12. In summary, we have shown by cytogenetics, segregant analysis, biochemical assay, and Southern blot analysis that human chromosome 1, most likely in the region 1 cen-1q12, corrects the defect in ADSS-deficient mutant Ade-H cells.

Original languageEnglish (US)
Pages (from-to)322-328
Number of pages7
JournalGenomics
Volume9
Issue number2
DOIs
StatePublished - 1991
Externally publishedYes

Fingerprint

Adenylosuccinate Synthase
Chromosomes, Human, Pair 1
Cricetulus
Adenine
Ovary
Human Chromosomes
Enzymes
Genes
Southern Blotting
Growth
Inosine Monophosphate
Cytogenetic Analysis
Adenosine Monophosphate
Chromosomes
purine

ASJC Scopus subject areas

  • Genetics

Cite this

@article{babccb70c9bc47208e88bb074cfa9a22,
title = "A gene correcting the defect in the CHO mutant Ade-H, deficient in a branch point enzyme (adenylosuccinate synthetase) of de novo purine biosynthesis, is located on the long arm of chromosome 1",
abstract = "Somatic hybrids between human cells and the Chinese hamster ovary (CHO) K1 mutant, Ade-H cells, were selected for purine prototrophy by growth in adenine-free medium. The Ade-H mutant is defective in the enzyme adenylosuccinate (AMPS) synthetase (ADSS; EC 6.3.4.4), which carries out the first of a two-step sequence in the biosynthesis of AMP from IMP, and therefore requires exogenous adenine for growth. The presence of the long arm of human chromosome 1 in the hybrids is 100{\%} concordant for the ability to growth in adenine-free medium and restoration of the enzyme activity. Hybrid segregants that lose the ability to grow in adenine-free medium lose all or a portion of chromosome 1 and enzyme activity. Southern blot hybridization with a chromosome 1-specific probe, BCMI, confirms the existence of human chromosome 1 in these hybrids. Analysis of a human/CHO translocation chromosome that arose in one of the hybrids suggests that the gene correcting the defect lies in the region 1 cen-1q12. In summary, we have shown by cytogenetics, segregant analysis, biochemical assay, and Southern blot analysis that human chromosome 1, most likely in the region 1 cen-1q12, corrects the defect in ADSS-deficient mutant Ade-H cells.",
author = "Li-Wen Lai and Hart, {Iris M.} and David Patterson",
year = "1991",
doi = "10.1016/0888-7543(91)90260-L",
language = "English (US)",
volume = "9",
pages = "322--328",
journal = "Genomics",
issn = "0888-7543",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - A gene correcting the defect in the CHO mutant Ade-H, deficient in a branch point enzyme (adenylosuccinate synthetase) of de novo purine biosynthesis, is located on the long arm of chromosome 1

AU - Lai, Li-Wen

AU - Hart, Iris M.

AU - Patterson, David

PY - 1991

Y1 - 1991

N2 - Somatic hybrids between human cells and the Chinese hamster ovary (CHO) K1 mutant, Ade-H cells, were selected for purine prototrophy by growth in adenine-free medium. The Ade-H mutant is defective in the enzyme adenylosuccinate (AMPS) synthetase (ADSS; EC 6.3.4.4), which carries out the first of a two-step sequence in the biosynthesis of AMP from IMP, and therefore requires exogenous adenine for growth. The presence of the long arm of human chromosome 1 in the hybrids is 100% concordant for the ability to growth in adenine-free medium and restoration of the enzyme activity. Hybrid segregants that lose the ability to grow in adenine-free medium lose all or a portion of chromosome 1 and enzyme activity. Southern blot hybridization with a chromosome 1-specific probe, BCMI, confirms the existence of human chromosome 1 in these hybrids. Analysis of a human/CHO translocation chromosome that arose in one of the hybrids suggests that the gene correcting the defect lies in the region 1 cen-1q12. In summary, we have shown by cytogenetics, segregant analysis, biochemical assay, and Southern blot analysis that human chromosome 1, most likely in the region 1 cen-1q12, corrects the defect in ADSS-deficient mutant Ade-H cells.

AB - Somatic hybrids between human cells and the Chinese hamster ovary (CHO) K1 mutant, Ade-H cells, were selected for purine prototrophy by growth in adenine-free medium. The Ade-H mutant is defective in the enzyme adenylosuccinate (AMPS) synthetase (ADSS; EC 6.3.4.4), which carries out the first of a two-step sequence in the biosynthesis of AMP from IMP, and therefore requires exogenous adenine for growth. The presence of the long arm of human chromosome 1 in the hybrids is 100% concordant for the ability to growth in adenine-free medium and restoration of the enzyme activity. Hybrid segregants that lose the ability to grow in adenine-free medium lose all or a portion of chromosome 1 and enzyme activity. Southern blot hybridization with a chromosome 1-specific probe, BCMI, confirms the existence of human chromosome 1 in these hybrids. Analysis of a human/CHO translocation chromosome that arose in one of the hybrids suggests that the gene correcting the defect lies in the region 1 cen-1q12. In summary, we have shown by cytogenetics, segregant analysis, biochemical assay, and Southern blot analysis that human chromosome 1, most likely in the region 1 cen-1q12, corrects the defect in ADSS-deficient mutant Ade-H cells.

UR - http://www.scopus.com/inward/record.url?scp=0025978788&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025978788&partnerID=8YFLogxK

U2 - 10.1016/0888-7543(91)90260-L

DO - 10.1016/0888-7543(91)90260-L

M3 - Article

C2 - 2004783

AN - SCOPUS:0025978788

VL - 9

SP - 322

EP - 328

JO - Genomics

JF - Genomics

SN - 0888-7543

IS - 2

ER -