Identification and Order of Sequential Mutations in Beta-actin Genes Isolated from Increasingly Tumorigenic Human Fibroblast Strains
Overview
Affiliations
We have sequenced the mutant beta-actin gene of a tumorigenic human fibroblast cell line (HuT-14T) and found that it carries three mutations that alter the amino acids at positions 36, 83, and 244 as well as a 22-base-pair "insertion" sequence, in the 5' intron, not present in a wild-type gene. The less tumorigenic cell line HuT-14, a progenitor of HuT-14T, has the same codon-244 mutation and the insertion sequence but not the other two mutations. A nontumorigenic cell line that is related to HuT-14 but that has no beta-actin mutations does carry the intron-length polymorphism. We conclude that the mutation at codon 244 occurred first in a beta-actin allele already bearing the 22-base-pair intron insert and that mutations at codons 36 and 83 arose subsequently during the selection for the HuT-14T phenotype. Rat-2 cells synthesize the appropriate charge-variant species of mutant actin protein when transfected with either the singly or the triply mutated beta-actin gene.
A method for rapidly screening functionality of actin mutants and tagged actins.
Rommelaere H, Waterschoot D, Neirynck K, Vandekerckhove J, Ampe C Biol Proced Online. 2004; 6:235-249.
PMID: 15514698 PMC: 524212. DOI: 10.1251/bpo94.
Modulation of cathepsin D activity in retinal pigment epithelial cells.
Rakoczy P, Lai C, Baines M, Di Grandi S, Fitton J, Constable I Biochem J. 1997; 324 ( Pt 3):935-40.
PMID: 9210419 PMC: 1218511. DOI: 10.1042/bj3240935.
Tropomyosin isoform switching in tumorigenic human fibroblasts.
Leavitt J, Latter G, Lutomski L, Goldstein D, Burbeck S Mol Cell Biol. 1986; 6(7):2721-6.
PMID: 3785208 PMC: 367830. DOI: 10.1128/mcb.6.7.2721-2726.1986.
Expression of transfected mutant beta-actin genes: transitions toward the stable tumorigenic state.
Leavitt J, Ng S, Varma M, Latter G, Burbeck S, Gunning P Mol Cell Biol. 1987; 7(7):2467-76.
PMID: 3614199 PMC: 365379. DOI: 10.1128/mcb.7.7.2467-2476.1987.
Leavitt J, Ng S, Aebi U, Varma M, Latter G, Burbeck S Mol Cell Biol. 1987; 7(7):2457-66.
PMID: 3614198 PMC: 365378. DOI: 10.1128/mcb.7.7.2457-2466.1987.