Hip3 interacts with the hira proteins Hip1 and Slm9 and is required for transcriptional silencing and accurate chromosome segregation

doi:10.1074/jbc.M512170200

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Papers In Press, published online ahead of print January 22, 2006
J. Biol. Chem, 10.1074/jbc.M512170200
Submitted on November 11, 2005
Revised on December 30, 2005
Accepted on January 22, 2006

Hip3 interacts with the hira proteins Hip1 and Slm9 and is required for transcriptional silencing and accurate chromosome segregation

Amanda Greenall, Emma S. Williams, Katherine A. Martin, Jeremy M. Palmer, Joe Gray, Cong Liu, and Simon K. Whitehall

Cell and Molecular Biosciences, University of Newcastle, The Medical School, Newcastle upon Tyne NE2 4HH

Corresponding Author: s.k.whitehall{at}ncl.ac.uk

The fission yeast HIRA proteins Hip1 and Slm9 are members of an evolutionarily conserved family of histone chaperones that are implicated in nucleosome assembly. Here we have used single-step affinity purification and MALDI-TOF mass spectrometry to identify factors that interact with both Hip1 and Slm9. This analysis identified Hip3, a previously uncharacterised 187 kDa protein, with similarity to S. cerevisiae Hir3. Consistent with this, cells disrupted for hip3+ exhibit a range of growth defects that are similar to those associated with loss of Hip1 and Slm9. These include temperature sensitivity, a cell cycle delay and synthetic lethality with cdc25-22. Furthermore, genetic analysis also indicates that disruption of hip3+ is epistatic with mutation of hip1+ and slm9+. Mutation of hip3+ alleviates transcriptional silencing at several heterochromatic loci including in the outer (otr) centromeric repeats, indicating that Hip3 is required for the integrity of pericentric heterochromatin. As a result loss of Hip3 function leads to high levels of minichromosome loss and an increased frequency of lagging chromosomes during mitosis. Importantly, the function of Hip1, Slm9 and Hip3 is not restricted to constitutive heterochromatic loci as these proteins also repress the expression of a number of genes including the Tf2 retrotransposons.

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