Gas2, a Growth Arrest-Specific Protein, Is a Component of the Microfilament Network System

• Published in: The Journal of cell biology 1992-06, Vol.117 (6), p.1251-1261
• Main Authors: Brancolini, Claudio, Bottega, Stefano, Schneider, Claudio
• Format: Article
• Language: English
• Subjects: 3T3 Cells; Actins; Analytical, structural and metabolic biochemistry; Animals; Antibodies; Bacteria; Biological and medical sciences; Blotting, Western; Cell Division; Cell growth; Cell lines; Cell Transformation, Neoplastic; Cells; Cellular biology; Cultured cells; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; G1 Phase; General aspects, investigation methods; Glutathione Transferase - genetics; Glutathione Transferase - metabolism; Growth Inhibitors - genetics; Growth Inhibitors - metabolism; Humans; Immunoprecipitation; Membrane Proteins - genetics; Membrane Proteins - metabolism; Mice; Microfilament Proteins - genetics; Microfilament Proteins - metabolism; Microfilaments; Microinjections; Microscopy, Fluorescence; NIH 3T3 cells; Precipitin Tests; Protein Processing, Post-Translational; Proteins; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Resting Phase, Cell Cycle
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.117.6.1251

In this report we analyze the protein product of a growth arrest-specific gene, gas2, by means of an affinity-purified antibody raised against the protein produced in bacteria. The regulation of Gas2 biosynthesis reflects the pattern of mRNA expression (Schneider, C., R. King, and L. Philipson. 1988. Cell. 54:787-793): its relative level is tightly associated with growth arrest. Gas2 seems to be regulated also at the posttranslational level via a phosphorylation mechanism. Gas2 is well conserved during the evolution with the same apparent molecular mass (36 kD) between mouse and human. We also demonstrate that Gas2 is a component of the microfilament system. It colocalizes with actin fiber, at the cell border and also along the stress fiber, in growth-arrested NIH 3T3 cells. The pattern of distribution, detected in arrested cells, can also be observed in growing cells when they are microinjected with the purified GST-Gas2 protein. In none of the analyzed oncogene-transformed NIH 3T3 cell lines was Gas2 expression induced under serum starvation.