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Vol. 15, No. 2, pp. 173-187, January 15, 2001
1 Program in Molecular Biology and Biotechnology,
2 Lineberger Comprehensive Cancer Center,
3 Department of Biochemistry and Biophysics,
4 Department of Biology, and 5 Curriculum in Genetics
and Molecular Biology, University of North Carolina, Chapel Hill, North
Carolina 27599, USA
Replication-associated histone genes encode the only metazoan mRNAs
that lack polyA tails, ending instead in a conserved 26-nt sequence
that forms a stem-loop. Most of the regulation of mammalian histone
mRNA is posttranscriptional and mediated by this unique 3' end.
Stem-loop-binding protein (SLBP) binds to the histone mRNA 3' end
and is thought to participate in all aspects of histone mRNA
metabolism, including cell cycle regulation. To examine SLBP function
genetically, we have cloned the gene encoding Drosophila SLBP
(dSLBP) by a yeast three-hybrid method and have isolated mutations in
dSLBP. dSLBP function is required both zygotically and
maternally. Strong dSLBP alleles cause zygotic lethality late in development and result in production of stable histone mRNA that
accumulates in nonreplicating cells. These histone mRNAs are
cytoplasmic and have polyadenylated 3' ends like other polymerase II transcripts. Hypomorphic dSLBP alleles support zygotic
development but cause female sterility. Eggs from these females contain
dramatically reduced levels of histone mRNA, and mutant embryos are not
able to complete the syncytial embryonic cycles. This is in part
because of a failure of chromosome condensation at mitosis that blocks normal anaphase. These data demonstrate that dSLBP is required in vivo
for 3' end processing of histone pre-mRNA, and that this is an
essential function for development. Moreover, dSLBP-dependent processing plays an important role in coupling histone mRNA production with the cell cycle.
[Key Words: Drosophila; cell cycle; SLBP; histone; replication; RNA processing]
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