Co-amplification of rRNA genes with CAD genes in N-(phosphonacetyl)-L-aspartate-resistant Syrian hamster cells

Mol Cell Biol. 1983 Nov;3(11):2066-75. doi: 10.1128/mcb.3.11.2066.

Abstract

The amplified CAD genes in N-(phosphonacetyl)-L-aspartate (PALA)-resistant Syrian hamster cells are located in an expanded chromosomal region emanating from the site of the wild-type gene at the tip of the short arm of chromosome B-9. The terminus of B-9 in PALA-sensitive cells contains a cluster of rRNA genes (i.e., a nucleolus organizer, rDNA). We have used a molecular clone containing sequences complementary to Syrian hamster 28S rRNA to investigate whether rDNA is coamplified with CAD genes in the PALA-resistant mutants. In situ hybridization of this probe to metaphase chromosomes demonstrates that rDNA and CAD genes do coamplify in two independently isolated PALA-resistant mutants. The tight linkage of CAD and rDNA genes was demonstrated by their coordinate translocation from B-9 to the end of the long arm of chromosome C-11 in one mutant. Blot hybridization studies substantiate the in situ hybridization results. Both types of analysis indicate that only one or two rDNA genes, on the average, are coamplified per CAD gene. The data are consistent with the model that unequal exchanges between rDNA genes mediate the amplification of CAD genes in the Syrian hamster mutants that were analyzed.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Aspartate Carbamoyltransferase*
  • Aspartic Acid / analogs & derivatives*
  • Aspartic Acid / pharmacology
  • Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)*
  • Cell Line
  • Chromosomes
  • Cricetinae
  • DNA Restriction Enzymes
  • Dihydroorotase*
  • Drug Resistance
  • Gene Amplification*
  • Genes
  • Mesocricetus
  • Models, Genetic
  • Multienzyme Complexes*
  • Nucleic Acid Hybridization
  • Organophosphorus Compounds / pharmacology*
  • Phosphonoacetic Acid / analogs & derivatives
  • Phosphonoacetic Acid / pharmacology*
  • Proteins / genetics*
  • RNA, Ribosomal / genetics*
  • Sister Chromatid Exchange

Substances

  • CAD trifunctional enzyme
  • Multienzyme Complexes
  • Organophosphorus Compounds
  • Proteins
  • RNA, Ribosomal
  • Aspartic Acid
  • sparfosic acid
  • Aspartate Carbamoyltransferase
  • DNA Restriction Enzymes
  • Dihydroorotase
  • Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)
  • Phosphonoacetic Acid