Nuclear actin and histones from the myxomycete Physarum flavicomum.
A method was established for the isolation and purification of nuclei in high yield from the microplasmodia of Physarum flavicomum. Purified nuclei was resistant to breakage by methods commonly employed for isolated plant and animal nuclei. Incubation of nuclei with 5 mM dithiothreitol at pH 9.2 was found to be the simplest and most effective method for breaking the nuclei. Several methods for the extraction of nuclear protein were compared. Incubation of nuclear lysates with either 2 M NaCl, with or without 5 M urea, or 1 M CaCl2 resulted in the extraction of nuclear actin together with histones. The histones were chemically fractionated into the five basic groups common to other eucaryotic tissue. Amino acid analyses of the total histone were also performed. Nuclear actin was found to have a molecular weight of 41,000 +/- 4,000 daltons as determined by SDS polyacrylamide gel electrophoresis. The amino acid composition of the nuclear actin was established.
D000199:Actins / Q000032:analysis* / Q000302:isolation & purification; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000032:analysis; D002843:Chromatin / Q000032:analysis; D006657:Histones / Q000032:analysis* / Q000302:isolation & purification; D010804:Physarum / Q000032:analysis* / Q000648:ultrastructure
D000199:Actins; D002843:Chromatin; D006657:Histones