Identification and functional analysis of PCNA1 and PCNA-like1 genes of Phaseolus coccineus.
Proliferating cell nuclear antigen (PCNA) is an essential factor in DNA replication and in many other processes in eukaryotic cells. Genetic analysis of Phaseolus coccineus showed the presence of at least two PCNA-like genes in the runner bean genome. Two PCNA genes have previously been found in a few plant species including Arabidopsis, tobacco, and maize. In these species, genes were nearly identical. Two cDNAs of P. coccineus PCNA (PcPCNA1 and PcPCNA-like1) have been identified that differ distinctly from each other. Interestingly, both the genetic organization of PcPCNA1 and PcPCNA-like1 genes and their expression patterns were similar, but these were the only similarities between these genes and their products. The identity between PcPCNA1 and PcPCNA-like1 at the amino acid level was only 54%, with PcPCNA-like1 lacking motifs that are crucial for the activity typical of PCNA. Consequently, these two proteins showed different properties. PcPCNA1 behaved like a typical PCNA protein: it formed a homotrimer and stimulated the activity of human DNA polymerase delta. In addition, PcPCNA1 interacted with a p21 peptide and was recognized by an anti-human PCNA monoclonal antibody PC10. By contrast, PcPCNA-like1 was detected as a monomer and was unable to stimulate the DNA polymerase delta activity. PcPCNA-like1 also could not interact with p21 and was not recognized by the PC10 antibody. Our results suggest that PcPCNA-like1 either is unable to function alone and therefore might be a component of the heterotrimeric PCNA ring or may have other, yet unknown functions. Alternatively, the PcPCNA-like1 gene may represent a pseudogene.
Strzalka
Wojciech
W
Department of Molecular Genetics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland.
Kaczmarek
Anna
A
Naganowska
Barbara
B
Ziemienowicz
Alicja
A
D000595:Amino Acid Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D032461:Chromosomes, Plant / Q000378:metabolism; D003001:Cloning, Molecular; D004258:DNA Polymerase III / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D000939:Epitopes / Q000737:chemistry; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics*; D018745:Genome, Plant / Q000235:genetics; D008677:Metaphase; D008969:Molecular Sequence Data; D027805:Phaseolus / Q000201:enzymology / Q000235:genetics*; D010802:Phylogeny; D020401:Primed In Situ Labeling; D018809:Proliferating Cell Nuclear Antigen / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018076:DNA, Complementary; D000939:Epitopes; D018809:Proliferating Cell Nuclear Antigen; D011994:Recombinant Proteins; D004258:DNA Polymerase III