Lens fiber differentiation correlated with activation of two different DNAases in lens embryonic cells.
In order to identify the different DNAases present in the lens differentiating tissue, we have used an assay which reveals their activity directly on DNA-containing gels after SDS polyacrylamide gel electrophoresis. DNAase renaturation from nuclear embryonic lens extracts does not occur after separation in 0.1% SDS polyacrylamide gel electrophoresis in contrast to that observed with purified micrococcal nuclease. When the SDS concentration in the running buffer and separating gel is decreased to 0.075%, renaturation of lens DNAase and enzyme activities are observed. Isoelectrofocusing was carried out in a polyacrylamide gel which was overlaid with an agarose gel containing DNA, permitting the visualization of the pI of DNAase activity. The presence of several DNAase isoenzymes was demonstrated in 11-day embryonic lenses. In epithelial lens nuclei, high molecular weight (MW) isoenzymes with basic pI were predominant. In post-mitotic fiber lens nuclei, two lower MW isoenzymes with acidic pI were detected as well as high MW activity with a basic pI.
INSERM U. 118, Unité de Recherches Gérontologiques, CNRS U.A. 630, Association Claude-Bernard, Paris, France.
M F
MF
Counis
E
E
Chaudun
Y
Y
Courtois
B
B
Allinquant
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002467:Cell Nucleus / Q000201:enzymology; D002642:Chick Embryo; D003851:Deoxyribonucleases / Q000302:isolation & purification* / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007526:Isoelectric Point; D007904:Lens Cortex, Crystalline / Q000196:embryology* / Q000201:enzymology; D007908:Lens, Crystalline / Q000196:embryology*; D008274:Magnesium / Q000494:pharmacology; D008836:Micrococcal Nuclease / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology
D012967:Sodium Dodecyl Sulfate; D003851:Deoxyribonucleases; D008836:Micrococcal Nuclease; D008274:Magnesium; D002118:Calcium