Co-immunoprecipitation of putative proteins that interact with mosquito proliferating cell nuclear antigen.
We have sequenced cDNAs encoding proliferating cell nuclear antigen (PCNA) from Aedes albopictus cells and from Aedes aegypti mosquitoes. The mosquito cDNAs contained an open reading frame encoding a 260 amino acid protein with a calculated mass of 29.0 kDa and a pI of 4.46. There was a single amino acid difference between PCNA proteins from Ae. albopictus and Ae. aegypti. In An. gambiae, the PCNA homolog contained 260 residues, and the pcna gene was interrupted by a single 67 nucleotide intron in the betaC2 region of the protein. A phylogenetic comparison grouped known Dipteran PCNA sequences into two clusters, representing the Nematocera and the Cyclorrhapha. PCNA transcripts measured 1.1 kb, and were stable, as was PCNA protein. Mosquito PCNA was efficiently recognized by a commercially available mouse anti-PCNA monoclonal antibody, which coprecipitated 29 kDa and 35 kDa proteins from mosquito cells representing different growth states. These results support the feasibility of recovering mosquito cell cycle inhibitory proteins by virtue of their interaction with PCNA.
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Department of Entomology, University of Minnesota, St Paul, 55108, USA.
Gerenday
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Coley
K M
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Fallon
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AM
D000330:Aedes / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D006224:Cricetinae; D018076:DNA, Complementary; D029721:Drosophila Proteins / Q000737:chemistry; D015870:Gene Expression; D017344:Genes, Insect; D047468:Immunoprecipitation; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry; D010802:Phylogeny; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D029721:Drosophila Proteins; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; C103336:dap protein, Drosophila