Preferential amplification of rearranged sequences near amplified adenylate deaminase genes.
In a previous study of three independent families of mutants selected for overproduction of adenylate deaminase (AMPD), we were not able to isolate a cDNA probe for the gene and so could not demonstrate its amplification directly. In addition to overproduction of AMPD, four proteins of unknown function, designated W, X, Y1, and Y2, accumulated, and by using the corresponding cDNA probes, we demonstrated amplification of all four genes. In independent mutant clones, sometimes all and sometimes only a subset of these genes were amplified. Assuming that all five genes are linked, the pattern of their coamplification suggested a genetic map in which AMPD lies between W and Y1. We show here that a two-step chromosome walk joins the W and Y1 genes, that the AMPD gene is the only expressed sequence between them, and that its amplification is indeed responsible for overproduction of the AMPD protein. In the course of this work, we cloned and studied two novel joints which mark rearrangements on either side of the AMPD gene. Each joint was generated independently in a single first-step mutant at single or low copy number. Remarkably, each joint was amplified preferentially in every second- and third-step mutant derived from the first-step line in which it was originally present, suggesting that the two independent rearrangements each generated amplification-prone structures.
Debatisse
M
M
Laboratory of Molecular Biology, Imperial Cancer Research Fund, London, United Kingdom.
Saito
I
I
Buttin
G
G
Stark
G R
GR
D000659:AMP Deaminase / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D006224:Cricetinae; D004247:DNA / Q000235:genetics; D005784:Gene Amplification; D005796:Genes; D008040:Genetic Linkage; D009709:Nucleotide Deaminases / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid
D004247:DNA; D009709:Nucleotide Deaminases; D000659:AMP Deaminase