The effects of mono-ADP-ribosylation on desmin assembly-disassembly.

Previous studies have shown that desmin, the muscle-specific intermediate filament protein, is a substrate for the endogenous muscle arginine-specific mono-ADP-ribosyltransferase and that ADP-ribosylation inhibits assembly of desmin into intermediate filaments (Huang et al., Exp. Cell Res. 226, 147-153, 1996). In this paper, the effects of mono-ADP-ribosylation on assembly and disassembly of desmin intermediate filaments were further characterized. First, it was found that ADP-ribosylated desmin does not coassemble with unmodified desmin and has no effect on assembly of unmodified desmin. Second, incubation of assembled desmin filaments with mono-ADP-ribosyltransferase and NAD+ results in disassembly of the filaments. Finally, the structural components of the attached ADP-ribose moiety responsible for altering the assembly of desmin into filaments were investigated by a stepwise cleavage of ADP-ribose with snake venom phosphodiesterase and alkaline phophatase, followed by analysis of assembly. The reactions catalyzed by these two enzymes were established using a desmin peptide as a substrate. Our results show that ribosylated desmin, but not phosphoribosylated desmin, was able to self-assemble into intermediate filaments, suggesting that the presence of a phosphate group is needed to alter desmin's assembly ability.