Microinjection of fos-specific antibodies blocks DNA synthesis in fibroblast cells

Riabowol, K. T., Vosatka, R. J., Ziff, E. B., Lamb, N. J., Feramisco, J. R. (1988) Microinjection of fos-specific antibodies blocks DNA synthesis in fibroblast cells. Molecular & Cellular Biology, 8 (4). pp. 1670-6. ISSN 0270-7306

URL: http://www.ncbi.nlm.nih.gov/pubmed/2454395
DOI: 10.1128/MCB.8.4.1670

Abstract

Transcription of the protooncogene c-fos is increased greater than 10-fold within minutes of treatment of fibroblasts with serum or purified growth factors. Recent experiments with mouse 3T3 cell lines containing inducible fos antisense RNA constructs have shown that induced fos antisense RNA transcripts cause either a marked inhibition of growth in continuously proliferating cells or, conversely, a minimal effect except during the transition from a quiescent (G0) state into the cell cycle. Since intracellular production of large amounts of antisense RNA does not completely block gene expression, we microinjected affinity-purified antibodies raised against fos to determine whether and when during the cell cycle c-fos expression was required for cell proliferation. Using this independent method, we found that microinjected fos antibodies efficiently blocked serum-stimulated DNA synthesis when injected up to 6 to 8 h after serum stimulation of quiescent REF-52 fibroblasts. Furthermore, when fos antibodies were injected into asynchronously growing cells, a consistently greater number of cells was prevented from synthesizing DNA than when cells were injected with nonspecific immunoglobulins. Thus, whereas the activity of c-fos may be necessary for transition of fibroblasts from G0 to G1 of the cell cycle, its function is also required during the early G1 portion of the cell cycle to allow subsequent DNA synthesis.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > antibodies
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > fibroblasts
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > fibroblasts
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > fibroblasts

bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > transcription factor
CSHL Authors:
Communities: CSHL labs
Depositing User: Gail Sherman
Date Deposited: 20 Sep 2017 19:04
Last Modified: 20 Sep 2017 19:04
PMCID: PMC363327
Related URLs:
URI: http://repository.cshl.edu/id/eprint/35207

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