title: The glycogen phosphorylase-2 promoter binding protein in Dictyostelium is replication protein A.
abstract: During Dictyostelium development, glycogen degradation is a crucial event that provides glucose monomers used in the synthesis of the essential structural components for cellular differentiation. The product of the developmentally regulated glycogen phosphorylase-2 gene (gp2) catalyzes the degradation. DNA-binding proteins were found to bind to a regulatory site of the gp2 gene in a stage-dependent pattern. Gel-shift analysis of undifferentiated amoebae cell extract revealed a protein migrating at 0.40 Rf, while 17 hour differentiated cell extract produced a species migrating at 0.32 Rf. Both the 0.32 and 0.40 Rf proteins were purified and found to consist of three subunits of 18, 35 and 62 kDa (for 0.40 Rf) or 81 kDa (for 0.32 Rf). Data base searches identified the protein as the Dictyostelium homologue of replication protein A (DdRPA). Amino acid sequence analysis showed identity between the 62 and 81 kDa subunits. Incubation of cell-free extracts under appropriate conditions at low pH, resulted in conversion of the 81 kDa to the 62 kDa subunit. Northern blot analysis revealed that the levels of expression of the large subunit of DdRPA were constant throughout differentiation and the size of the mRNA was the same at all stages of development. The results raise the possibility that pH induced post-translational modifications of DdRPA are involved in events that halt cell proliferation and induce differentiation in Dictyostelium.
authors.affiliation: Biology Department, Molecular and Cellular Biology Section, Virginia Polytechnic Institute and State University, Blacksburg, VA, 24061, USA.
authors.forename: X
authors.initials: X
authors.lastname: Wen
authors.forename: P
authors.initials: P
authors.lastname: Khampang
authors.forename: C L
authors.initials: CL
authors.lastname: Rutherford
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002455:Cell Division; D003001:Cloning, Molecular; D004261:DNA Replication; D016054:DNA, Protozoan / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004023:Dictyostelium / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D017125:Genes, Protozoan; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D008970:Molecular Weight; D006005:Phosphorylases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D011487:Protein Conformation; D015800:Protozoan Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D051716:Replication Protein A; D017386:Sequence Homology, Amino Acid
chemical_list: D016054:DNA, Protozoan; D004268:DNA-Binding Proteins; D015800:Protozoan Proteins; C497167:RPA1 protein, human; D051716:Replication Protein A; D006005:Phosphorylases; C105247:glycogen phosphorylase 2 protein, Dictyostelium discoideum