Journal of Experimental Botany, Vol 50, 1315-1322, Copyright © 1999 by Oxford University Press
P Sabelli, J Parker and P Barlow
A partial cDNA from maize, ROA, encoding a protein
homologous to the MCM3 family of essential factors for the initiation of
DNA replication, has been isolated previously. In the present work, a
longer version of the original ROA cDNA, encoding a
full-length protein, was isolated and termed ZmROA1.
In addition, three other closely related cDNAs, ZmROA2,
ZmROA3 and ZmROA4, were also isolated.
ZmROA2 end ZmROA3 appear to
encode full-length proteins, whereas ZmROA4 a partial
polypeptide. Two clusters of basic amino acids comprising putative nuclear
localization signals were identified in the N-terminal domain of these
proteins, together with a potential leucine zipper. Immunofluorescence
studies on cycling meristematic root-tip cells revealed that these proteins
are localized in the nucleus throughout interphase with a pattern
overlapping that of chromatin. However, as chromatin condenses at prophase,
ZmROA proteins become increasingly distinct from chromatin and appear
totally dissociated from the segregating chromosomes during mitosis. This
behaviour is consistent with a role in restricting DNA replication to only
one round in each cell cycle. A 4.7 kb genomic sequence was also isolated,
comprising part of the ZmROA2 gene, with approximately
3.2 kb of promoter sequence, and 600 bp of 5' sequence of the
ZmROA2 cDNA, including the first four exons and three
introns. Several putative regulatory elements were identified in the
promoter sequence. This is the first report on promoter and cDNA sequences
encoding full-length MCM3 homologues from higher plants and the
distribution of such proteins during the plant cell cycle.Key
words: Cell cycle, DNA replication licensing factor, MCM
proteins, plant.
ARTICLES
cDNA and promoter sequences for MCM3 homologues from maize, and protein localization in cycling cells
Institute of Molecular and Cell Biology, National University of Singapore, 30 Medical Drive, Singapore 117609, Republic of Singapore; IACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, Long Ashton, Bristol BS18 9AF, UK; Corresponding author at: Via del Parco 50, 57128 Livorno, Italy. E-mail: paolosabelli@iol.it
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