title: Characterization of monomeric DNA-binding protein Histone H1 in Leishmania braziliensis.
abstract: Histone H1 in Leishmania presents relevant differences compared to higher eukaryote counterparts, such as the lack of a DNA-binding central globular domain. Despite that, it is apparently fully functional since its differential expression levels have been related to changes in chromatin condensation and infectivity, among other features. The localization and the aggregation state of L. braziliensis H1 has been determined by immunolocalization, mass spectrometry, cross-linking and electrophoretic mobility shift assays. Analysis of H1 sequences from the Leishmania Genome Database revealed that our protein is included in a very divergent group of histones H1 that is present only in L. braziliensis. An antibody raised against recombinant L. braziliensis H1 recognized specifically that protein by immunoblot in L. braziliensis extracts, but not in other Leishmania species, a consequence of the sequence divergences observed among Leishmania species. Mass spectrometry analysis and in vitro DNA-binding experiments have also proven that L. braziliensis H1 is monomeric in solution, but oligomerizes upon binding to DNA. Finally, despite the lack of a globular domain, L. braziliensis H1 is able to form complexes with DNA in vitro, with higher affinity for supercoiled compared to linear DNA.
authors.affiliation: Instituto de Enfermedades Tropicales y Salud Pública de Canarias. Universidad de La Laguna. Av. Astrofisico Fco. Sanchez, s/n. 38202 La Laguna, Tenerife, Spain. ecarmelo@ull.es
authors.forename: Emma
authors.initials: E
authors.lastname: Carmelo
authors.forename: Gloria
authors.initials: G
authors.lastname: González
authors.forename: Teresa
authors.initials: T
authors.lastname: Cruz
authors.forename: Antonio
authors.initials: A
authors.lastname: Osuna
authors.forename: Mariano
authors.initials: M
authors.lastname: Hernández
authors.forename: Basilio
authors.initials: B
authors.lastname: Valladares
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism; D015153:Blotting, Western; D004278:DNA, Superhelical / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D030541:Databases, Genetic; D024202:Electrophoretic Mobility Shift Assay; D006657:Histones / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D007120:Immunochemistry; D007892:Leishmania braziliensis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016773:Leishmaniasis, Cutaneous / Q000469:parasitology; D013058:Mass Spectrometry; D010802:Phylogeny; D058105:Polymerization; D054730:Protein Interaction Domains and Motifs / Q000235:genetics*; D011817:Rabbits; D012996:Solutions / Q000378:metabolism
chemical_list: D000906:Antibodies; D004278:DNA, Superhelical; D004268:DNA-Binding Proteins; D006657:Histones; D012996:Solutions