Pe. Hart et Sm. Wolniak, Molecular cloning of a centrin homolog from Marsilea vestita and evidence for its translational control during spermiogenesis, BIOC CELL B, 77(2), 1999, pp. 101-108
Citations number
48
Categorie Soggetti
Cell & Developmental Biology
Journal title
BIOCHEMISTRY AND CELL BIOLOGY-BIOCHIMIE ET BIOLOGIE CELLULAIRE
Spermiogenesis in the water fern Marsilea vestita is a process that reaches
completion 11 h after dry microspores are immersed in an aqueous medium at
20 degrees C. Each microspore produces 32 spermatozoids and each spermatoz
oid has a coiled cell body and approximately 140 cilia. The spermatids make
basal bodies de novo, from a structure known as a blepharoplast. From the
onset of development, the spores contain a large quantity of protein and st
ored mRNA. We have found previously that centrin, a protein involved in the
function of microtubule organizing centers and present in association with
basal bodies in motile cells, is made in large quantity approximately 4 h
after the microspores are placed into liquid medium. In this paper, we show
that a centrin cDNA (MvCen1) we isolated from M. vestita closely resembles
centrin cDNAs from other eukaryotic organisms, MvCen1, synthesized in Esch
erichia coli as a GST-fusion protein, reacted with anti-centrin monoclonal
antibodies on immunoblots. Northern blot analysis demonstrates that centrin
mRNA is present in the dry microspore at the time of imbibition, at levels
that remain constant over 10 h of development and are unaffected by treatm
ent of spores with alpha-amanitin. The centrin transcripts, stored in dry m
icrospores, cannot be translated in vitro for at least 30 min after imbibit
ion.