Cloning of murine gelsolin and its regulation during differentiation of embryonal carcinoma cells.
The regulation of gelsolin levels during differentiation of the murine embryonal carcinoma cell line, PC-13, was investigated using nucleic acid and immunological probes. A cDNA clone, Mu-319, which contained the entire coding sequence for the cytoplasmic form of murine gelsolin was isolated using a polyclonal antibody. Gelsolin was detected in several cell lines but was not detectable in three undifferentiated embryonal carcinoma cell lines. Levels of gelsolin mRNA increased 10-fold during the differentiation of the murine embryonal carcinoma cell line, PC-13. Differentiation of PC-13 was accompanied by changes in cell shape, from small indistinct cells to large flat cells. The accumulation of gelsolin mRNA in PC-13 cells began 12-24 h after addition of the differentiation-inducing agents. In comparison, 2-5A-dependent RNase activity showed a 40-fold increase beginning after 24 to 36 h and c-fos mRNA were shown to increase about 9-fold beginning 36 to 60 h after induction of differentiation. The levels of gelsolin per se, as determined by immunoreactivity were also shown to increase with differentiation of PC-13 cells. These results suggest that gelsolin may play a role in the restructuring of actin filaments which accompanies the dramatic changes in cell shape during differentiation.
Dieffenbach
C W
CW
Department of Pathology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799.
SenGupta
D N
DN
Krause
D
D
Sawzak
D
D
Silverman
R H
RH
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D002454:Cell Differentiation; D002460:Cell Line; D002482:Cellulose / Q000031:analogs & derivatives; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification; D054278:Embryonal Carcinoma Stem Cells; D018260:Gelsolin; D005786:Gene Expression Regulation; D051379:Mice; D008840:Microfilament Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D014411:Neoplastic Stem Cells / Q000378:metabolism* / Q000473:pathology; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012117:Resins, Synthetic; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000473:pathology
C045415:2',5'-oligoadenylate-cellulose; D002135:Calcium-Binding Proteins; D018260:Gelsolin; D008840:Microfilament Proteins; D011993:Recombinant Fusion Proteins; D012117:Resins, Synthetic; D002482:Cellulose; D004247:DNA