Regulatory role of arginine-specific mono(ADP-ribosyl)transferase in muscle cells.
Earlier we demonstrated that meta-iodobenzylguanidine (MIBG), a specific inhibitor of arginine mono-ADP-ribosylation blocks proliferation and differentiation of chick skeletal myogenic cells in culture (Exp. Cell Res., 1992, 201:33-42). Membrane fractions from 4-day, myotube cultures of embryonic chick muscle cells were incubated with 32P-NAD+. Several proteins were labeled, but labeling of two hands of about 53 and 36 kDa appeared to be due to arginyl ADP-ribosylation. Immunoprecipitation with D3 monoclonal antibody to the intermediate filament protein desmin, SDS-PAGE and autoradiography demonstrated that the 53 kDa band contained desmin, and that this desmin is ADP-ribosylated by the endogenous arginine-specific mono(ADP-ribosyl)transferase (Exp. Cell Res., 1996, in press). Desmin is the muscle-specific intermediate filament protein, and it appears to be one of the first muscle-specific proteins expressed during terminal myogenic differentiation. We have examined whether desmin can be ADP-ribosylated in muscle cells by use of polyclonal antibodies for ADP-ribosylated arginyl residues. We have found that soluble desmin is present in 5-6 day myogenic cell cultures and that this desmin contains ADP-ribose, demonstrating that desmin is ADP-ribosylated in skeletal muscle cells. We also found that purified avian desmin contains antigenic material that reacts with these antibodies. In both cases, NaCl had no effect on the reactivity, but NH2OH did, which is consistent with an arginine-ADPR linkage. In summary, these results suggest that ADP-ribosylation is an important regulatory mechanism in differentiating muscle cells, and that the intermediate filament protein desmin is an important substrate for modification in muscle cells.
Graves
D J
DJ
Department of Biochemistry and Biophysics, Iowa State University, Ames 50011, USA.
Huiatt
T W
TW
Zhou
H
H
Huang
H Y
HY
Sernett
S W
SW
Robson
R M
RM
McMahon
K K
KK
D036002:ADP Ribose Transferases / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D002642:Chick Embryo; D003893:Desmin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism*
D003893:Desmin; D036002:ADP Ribose Transferases