title: The Drosophila histone variant H2A.V works in concert with HP1 to promote kinetochore-driven microtubule formation.
abstract: Unlike other organisms that have evolved distinct H2A variants for different functions, Drosophila melanogaster has just one variant which is capable of filling many roles. This protein, H2A.V, combines the features of the conserved variants H2A.Z and H2A.X in transcriptional control/heterochromatin assembly and DNA damage response, respectively. Here we show that mutations in the gene encoding H2A.V affect chromatin compaction and perturb chromosome segregation in Drosophila mitotic cells. A microtubule (MT) regrowth assay after cold exposure revealed that loss of H2A.V impairs the formation of kinetochore-driven (k) fibers, which can account for defects in chromosome segregation. All defects are rescued by a transgene encoding H2A.V that lacks the H2A.X function in the DNA damage response, suggesting that the H2A.Z (but not H2A.X) functionality of H2A.V is required for chromosome segregation. We also found that loss of H2A.V weakens HP1 localization, specifically at the pericentric heterochromatin of metaphase chromosomes. Interestingly, loss of HP1 yielded not only telomeric fusions but also mitotic defects similar to those seen in H2A.V null mutants, suggesting a role for HP1 in chromosome segregation. We also show that H2A.V precipitates HP1 from larval brain extracts indicating that both proteins are part of the same complex. Moreover, we found that the overexpression of HP1 rescues chromosome missegregation and defects in the kinetochore-driven k-fiber regrowth of H2A.V mutants indicating that both phenotypes are influenced by unbalanced levels of HP1. Collectively, our results suggest that H2A.V and HP1 work in concert to ensure kinetochore-driven MT growth.
authors.affiliation: a Dipartimento di Biologia e Biotecnologie "C. Darwin" ; Sapienza Università di Roma ; Roma , Italy.
authors.forename: Fiammetta
authors.initials: F
authors.lastname: Vernì
authors.forename: Giovanni
authors.initials: G
authors.lastname: Cenci
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002843:Chromatin / Q000502:physiology; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D020090:Chromosome Segregation / Q000502:physiology*; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism / Q000502:physiology*; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D018386:Kinetochores / Q000502:physiology*; D007814:Larva / Q000378:metabolism; D008870:Microtubules / Q000502:physiology*; D008938:Mitosis / Q000502:physiology*
chemical_list: D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D029721:Drosophila Proteins; D006657:Histones; C452193:heterochromatin protein 1, Drosophila
keywords: Drosophila; H2A.V; HP1; chromosome segregation; mitosis