Immunoelectron microscopic analysis of the intracellular distribution of primer recognition proteins, annexin 2 and phosphoglycerate kinase, in normal and transformed cells.
Primer recognition proteins (PRP) are accessory proteins for DNA polymerase alpha in lagging strand DNA replication. We have previously reported that the PRP consist of a complex of two proteins identified as 3-phosphoglycerate kinase (PGK) and the protein-tyrosine kinase substrate, annexin 2 monomer. The physiological role of annexin 2 is not known. Two pools of annexin 2 exist in cells. A majority of annexin 2 is localized with the plasma membrane as a heterotetramer in association with a light chain. Monomer annexin 2 is cytosolic. The identification of annexin 2 monomer as a part of the PRP complex represents one of the physiological roles of this protein in cells. To function as PRP, annexin 2 and PGK would have to be present in the cell nucleus. To investigate whether monomer annexin 2 is indeed associated with nuclear DNA synthesis, we investigated the presence of annexin 2 and PGK in the cell nucleus. In this paper, we demonstrate the presence of annexin 2 and PGK in nuclear extracts. The nuclear fraction of these proteins represents a small subset of the total cellular pools. Immunoelectron-microscopic analyses using anti-PRP antisera demonstrate the distribution of these proteins in HeLa cell nuclei and cytoplasm. Under identical conditions, an anti-cytokeratin monoclonal antibody preferentially labels the plasma membrane without detectable intracellular staining. The distribution of annexin 2 and PGK in both nuclei and cytoplasm is similarly observed in cells from normal tissues such as freshly isolated rat hepatocytes and hamster pancreatic tissue.(ABSTRACT TRUNCATED AT 250 WORDS)
Kumble
K D
KD
Department of Biochemistry, University of Nebraska Medical Center, Omaha 68198-4525.
Vishwanatha
J K
JK
D000818:Animals; D017302:Annexins; D002135:Calcium-Binding Proteins / Q000032:analysis*; D002453:Cell Cycle; D002462:Cell Membrane / Q000737:chemistry; D002467:Cell Nucleus / Q000737:chemistry* / Q000201:enzymology; D002471:Cell Transformation, Neoplastic; D006224:Cricetinae; D003593:Cytoplasm / Q000737:chemistry* / Q000201:enzymology; D004261:DNA Replication; D006367:HeLa Cells; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000648:ultrastructure; D008565:Membrane Proteins / Q000032:analysis*; D016253:Microscopy, Immunoelectron; D010179:Pancreas / Q000737:chemistry / Q000648:ultrastructure; D010735:Phosphoglycerate Kinase / Q000032:analysis*; D051381:Rats
D017302:Annexins; D002135:Calcium-Binding Proteins; D008565:Membrane Proteins; D010735:Phosphoglycerate Kinase