Phosphorylation of subunit proteins of intermediate filaments from chicken muscle and nonmuscle cells

O’Connor, Clare M.; Balzer, David R.; Lazarides, Elias

doi:10.1073/pnas.76.2.819

Cited by 102 publications

(69 citation statements)

References 23 publications

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“…This pattern of vimentin-derived peptides is identical to that described in extracts of vascular smooth muscle (9), skeletal muscle (24) and nonmuscle cells grown in vitro (6,8,23), and in cells of the early mammalian embryo (15).…”

Section: Discussionmentioning

confidence: 59%

Proteolysis of vimentin and desmin by the Ca2+-activated proteinase specific for these intermediate filament proteins.

Nelson¹,

Traub²

1983

Mol. Cell. Biol.

148

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The degradation of vimentin and desmin by the Ca2+-activated proteinase specific for these intermediate filament proteins proceeds in two stages in the form of a limited proteolysis. At first, the reaction is very rapid, with the stepwise and complete removal of a peptide (ca. 9,000 daltons) from the N-terminal of vimentin and desmin. This results in the production of a characteristic "staircase" of degradation products, as seen in two-dimensional polyacrylamide gel electrophoresis. The second stage of proteolysis is characterized by the accumulation of peptides which are resistant to further proteolysis; this is due not to product inhibition but to the fact that these peptides are not substrates for the proteinase and therefore do not protect the latter from inactivation (autodigestion). In vitro phosphorylation of the substrates does not affect proteinase activity, probably because the phosphorylation site is located towards the C-terminal of the molecules. The specific and limited proteolysis of vimentin and desmin results in the deletion of the nucleic acid binding and filament assembly site of these proteins, indicating that the Ca2'-activated proteinase plays a role in regulating the function(s) of these intermediate filament proteins, rather than their simple turnover during the cell cycle.Intermediate (10-nm) filaments are a heterogeneous class of protein fibrils which can be subdivided into five groups based on their subunit composition and cellular origin (1,2,7,17 helix (particularly at the amino terminal) interspersed with regions of non-a-helix (11,32,33). Most recently, it has been shown that there is ca. 70% amino acid sequence homology at the carboxy terminal between vimentin and desmin (11-13). In addition, both proteins bind to nucleic acids (38).Vimentin and desmin are also very susceptible to proteolysis in situ (17). This is due to the activity of a highly specific, Ca2'-activated, neutral proteinase for which we have recently described the purification and characterization (18,19). The proteinase appears to be coconserved during evolution with the substrate vimentin (20) and is present in reduced amounts in cells in which little or no vimentin is synthesized (36). Together with the fact that the Ca2+-activated proteinase does not significantly degrade other proteins (18,19), these results indicate that the proteinase plays an important role in the regulation of the function of vimentin and desmin. To investigate this, we have analyzed in detail the effect of the proteinase on vimentin and desmin. We show that there is only a limited proteolysis of a 9-kilodalton (kd) fragment from the N-terminal, resulting in the accumulation of large peptides which are resistant to further proteolysis. This accumulation is due not to product inhibition of proteolytic activity but to the fact that these peptides are poor substrates,

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Section: Discussionmentioning

confidence: 59%

Proteolysis of vimentin and desmin by the Ca2+-activated proteinase specific for these intermediate filament proteins.

Nelson¹,

Traub²

1983

Mol. Cell. Biol.

148

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“…Electron microscopic studies have demonstrated that filaments with a diameter of 10 (3,13). When muscle cell cytoskeletal proteins are electrophoresed on two-dimensional gels, both desmin and vimentin show microheterogeneity (8,13).…”

mentioning

confidence: 99%

“…When muscle cell cytoskeletal proteins are electrophoresed on two-dimensional gels, both desmin and vimentin show microheterogeneity (8,13). Desmin appears as a set of isoelectric variants with a molecular weight of 50,000, and vimentin appears as a series of variants with a molecular weight of 52,000.…”

mentioning

confidence: 99%

In vitro translation of the intermediate filament proteins desmin and vimentin.

O’Connor¹,

Asai²,

Flytzanis³

et al. 1981

Mol. Cell. Biol.

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“…In uninfected cells, the labelled vimentin had the same mobility as the marker, and its mobility relative to other major cellular proteins (e.g. actin) was similar to that previously reported (Brown et al, 1976;Ben-Zeev et al, 1979;Cabral & Gottesman, 1979;O'Connor et al, 1979). Finally, the identity of vimentin was further established by immunoprecipitation with anti-vimentin antibodies [a kind gift of Dr R. Hynes (Hynes & Destree, 1978)].…”

Section: Qualitative Change In Vimentin Of Fv3-infected Bhk Cellsmentioning

confidence: 67%

Interaction of Frog Virus 3 with the Cytomatrix. IV. Phosphorylation of Vimentin Precedes the Reorganization of Intermediate Filaments around the Virus Assembly Sites

Chen

Goorha

Murti

1986

Journal of General Virology

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SUMMARYFrog virus 3 (FV3) assembles in morphologically distinct assembly sites in the cytoplasm of infected cells. As the assembly sites form, the intermediate filaments (IF) aggregate, delimit the assembly sites, and remain so throughout infection. To determine the molecular basis of reorganization of IF, we analysed the vimentin of uninfected and FV3-infected cells by two-dimensional gel electrophoresis. The results showed that (i) the vimentin was more acidic in FV3-infected cells than in uninfected cells, (ii) the acidification of vimentin in FV3-infected cells was possibly due to a fourfold increase in phosphorylation, and (iii) the phosphorylation of vimentin preceded the reorganization of IF around virus assembly sites. A temperature-sensitive mutant of FV3 (ts9467), which at the non-permissive temperature neither reorganized IF nor formed assembly sites, failed to increase the phosphorylation of vimentin. Together, the above results suggest that changes in phosphorylation may modulate IF organization and that changes in IF organization are required for FV3 assembly site formation.

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Phosphorylation of subunit proteins of intermediate filaments from chicken muscle and nonmuscle cells

Cited by 102 publications

References 23 publications

Proteolysis of vimentin and desmin by the Ca2+-activated proteinase specific for these intermediate filament proteins.

Proteolysis of vimentin and desmin by the Ca2+-activated proteinase specific for these intermediate filament proteins.

In vitro translation of the intermediate filament proteins desmin and vimentin.

Interaction of Frog Virus 3 with the Cytomatrix. IV. Phosphorylation of Vimentin Precedes the Reorganization of Intermediate Filaments around the Virus Assembly Sites

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