Proliferating cell nuclear antigen from a basidiomycete, Coprinus cinereus. Alternative truncation and expression in meiosis.
The primary purpose of the present study was to investigate whether DNA replication at meiotic prophase also requires replication factors, especially proliferating cell nuclear antigen (PCNA). We cloned PCNA cDNAs (CoPCNA) from a cDNA library made from basidia of the basidiomycete, Coprinus cinereus. Interestingly, although CoPCNA is a single-copy gene in the genome, two different PCNA cDNA species were isolated using degenerate primers and a meiotic cDNA library, and were designated as CoPCNA-alpha and CoPCNA-beta. CoPCNA-beta was made by truncating at specific sites in CoPCNA-alpha mRNA, 5'-AAGAAGGAGAAG-3' and 5'-GAAGAGGAAGAA-3'. Both of these sequences were present in exon IV in the genomic sequence, and interestingly the former was the same as the inverse sequence of the latter. CoPCNA-alpha was 107 amino acids larger than human PCNA, and so the 107 amino-acid sequence was inserted in a loop, the so-called D2E2 loop, in human PCNA. Northern blotting analysis indicated that CoPCNA was expressed not only at premeiotic S but also at the meiotic prophase stages such as leptotene and early zygotene, just before and when karyogamy occurs and the homologous chromosomes pair. Western blotting analysis using anti-(CoPCNA-alpha) Ig revealed that at least two CoPCNA mRNAs before and after truncation were translated at the meiotic prophase as CoPCNA-alpha and CoPCNA-beta.
Hamada
Fumika
F
Department of Applied Biological Science, Faculty of Science and Technology, Science University of Tokyo, Japan.
Namekawa
Satoshi
S
Kasai
Nobuyuki
N
Nara
Takayuki
T
Kimura
Seisuke
S
Sugawara
Fumio
F
Sakaguchi
Kengo
K
D001487:Basidiomycota / Q000235:genetics*; D015152:Blotting, Northern; D015153:Blotting, Western; D003198:Computer Simulation; D018076:DNA, Complementary / Q000032:analysis / Q000302:isolation & purification; D008540:Meiosis; D008961:Models, Structural; D018809:Proliferating Cell Nuclear Antigen / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis
D018076:DNA, Complementary; D018809:Proliferating Cell Nuclear Antigen; D012333:RNA, Messenger