Disruption of the coordinate expression of muscle genes in a transfected BC3H1 myoblast cell line producing a low level of the adenovirus E1A transforming protein.
Mouse BC3H1 myoblasts were stably transfected with the adenovirus 5 E1A gene. One clonal line, BC3E7, was found to differ in some important respects from those previously reported for E1A-transformed myoblasts. In contrast to BC3H1 cells which differentiate when confluent in medium containing 0.5% fetal calf serum (FCS), BC3E7 cells failed to elongate and align, to express acetylcholine receptor and creatine kinase, and to down-regulate expression of beta- and gamma-actins and tropomyosin isoform (TM) 1. However, increased synthesis of TMs 2, 3, and 4, and myosin light chain 1 associated with differentiation in BC3H1 still occurred in BC3E7 cells, and most surprisingly, alpha-actin was produced at a significant level in both proliferating and confluent BC3E7 cells. Interestingly, myogenin was expressed in confluent BC3E7 cells in 0.5% FCS, but not in 20%. The level of E1A expression in BC3E7 cells was found to be very low by analysis of mRNA, by immunoprecipitation of E1A protein, and by the ability of BC3E7 cells to complement the E1A-deficient adenovirus mutant dl312. These results suggest that different levels of E1A may be needed to repress different promoters and that E1A does not block myogenic differentiation by repressing myogenin expression, but represses each muscle gene independently.
Mymryk
J S
JS
Department of Biochemistry, McMaster University, Hamilton, Ont., Canada.
Oakes
J D
JD
Muthuswamy
S K
SK
D'Amico
P
P
Bayley
S T
ST
Lee
R W
RW
D017500:Adenovirus E1A Proteins / Q000235:genetics / Q000502:physiology*; D000260:Adenoviruses, Human / Q000235:genetics / Q000502:physiology*; D000818:Animals; D002454:Cell Differentiation; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation / Q000502:physiology; D051379:Mice; D009124:Muscle Proteins / Q000096:biosynthesis* / Q000235:genetics; D009132:Muscles / Q000166:cytology; D018008:Myogenin; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D017500:Adenovirus E1A Proteins; D009124:Muscle Proteins; C503801:Myog protein, mouse; D018008:Myogenin; D011993:Recombinant Fusion Proteins