title: Functional and Structural Insights of the Zinc-Finger HIT protein family members Involved in Box C/D snoRNP Biogenesis.
abstract: Zf–HIT family members share the zf–HIT domain (ZHD), which is characterized by a fold in “treble-clef” through interleaved CCCC and CCHC ZnF motifs that both bind a zinc atom. Six proteins containing ZHD are present in human and three in yeast proteome, all belonging to multimodular RNA/protein complexes involved in gene regulation, chromatin remodeling, and snoRNP assembly. An interesting characteristic of the cellular complexes that ensure these functions is the presence of the RuvBL1/2/Rvb1/2 ATPases closely linked with zf–HIT proteins. Human ZNHIT6/BCD1 and its counterpart in yeast Bcd1p were previously characterized as assembly factors of the box C/D snoRNPs. Our data reveal that the ZHD of Bcd1p is necessary but not sufficient for yeast growth and that the motif has no direct RNA-binding capacity but helps Bcd1p maintain the box C/D snoRNAs level in steady state. However, we demonstrated that Bcd1p interacts nonspecifically with RNAs depending on their length. Interestingly, the ZHD of Bcd1p is functionally interchangeable with that of Hit1p, another box C/D snoRNP assembly factor belonging to the zf–HIT family. This prompted us to use NMR to solve the 3D structures of ZHD from yeast Bcd1p and Hit1p to highlight the structural similarity in the zf–HIT family. We identified structural features associated with the requirement of Hit1p and Bcd1p ZHD for cell growth and box C/D snoRNA stability under heat stress. Altogether, our data suggest an important role of ZHD could be to maintain functional folding to the rest of the protein, especially under heat stress conditions.
authors.affiliation: Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), UMR 7365, CNRS Université de Lorraine, Biopôle, Campus Biologie-Santé, 9 Avenue de la Forêt de Haye, CS 50184, 54505 Vandœuvre-lès-Nancy, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Bragantini
authors.affiliation: Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), UMR 7365, CNRS Université de Lorraine, Biopôle, Campus Biologie-Santé, 9 Avenue de la Forêt de Haye, CS 50184, 54505 Vandœuvre-lès-Nancy, France.
authors.forename: Decebal
authors.initials: D
authors.lastname: Tiotiu
authors.affiliation: Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), UMR 7365, CNRS Université de Lorraine, Biopôle, Campus Biologie-Santé, 9 Avenue de la Forêt de Haye, CS 50184, 54505 Vandœuvre-lès-Nancy, France.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Rothé
authors.affiliation: BioOrganic Mass Spectrometry Laboratory (LSMBO), IPHC, Université de Strasbourg, 25 rue Becquerel, 67087 Strasbourg, France; IPHC, UMR 7178, CNRS, 67087 Strasbourg, France.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Saliou
authors.affiliation: Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), UMR 7365, CNRS Université de Lorraine, Biopôle, Campus Biologie-Santé, 9 Avenue de la Forêt de Haye, CS 50184, 54505 Vandœuvre-lès-Nancy, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Marty
authors.affiliation: BioOrganic Mass Spectrometry Laboratory (LSMBO), IPHC, Université de Strasbourg, 25 rue Becquerel, 67087 Strasbourg, France; IPHC, UMR 7178, CNRS, 67087 Strasbourg, France.
authors.forename: Sarah
authors.initials: S
authors.lastname: Cianférani
authors.affiliation: Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), UMR 7365, CNRS Université de Lorraine, Biopôle, Campus Biologie-Santé, 9 Avenue de la Forêt de Haye, CS 50184, 54505 Vandœuvre-lès-Nancy, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Charpentier
authors.affiliation: FR 3209, CNRS-Université de Lorraine, Bioingénierie Moléculaire, Cellulaire et Thérapeutique, Biopôle, Campus Biologie-Santé, CS 50184, 54505 Vandœuvre-lès-Nancy Cedex, France. Electronic address: marc.quinternet@univ-lorraine.fr.
authors.forename: Marc
authors.initials: M
authors.lastname: Quinternet
authors.affiliation: Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), UMR 7365, CNRS Université de Lorraine, Biopôle, Campus Biologie-Santé, 9 Avenue de la Forêt de Haye, CS 50184, 54505 Vandœuvre-lès-Nancy, France. Electronic address: xavier.manival@univ-lorraine.fr.
authors.forename: Xavier
authors.initials: X
authors.lastname: Manival
mesh_terms: D006358:Hot Temperature; D000075723:Kruppel-Like Factor 6 / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D011487:Protein Conformation; D017510:Protein Folding; D055550:Protein Stability; D020718:Ribonucleoproteins, Small Nucleolar / Q000737:chemistry / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000254:growth & development* / Q000528:radiation effects; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D013312:Stress, Physiological; D016335:Zinc Fingers
chemical_list: C000594604:Hit1 protein, S cerevisiae; D000075723:Kruppel-Like Factor 6; D020718:Ribonucleoproteins, Small Nucleolar; D029701:Saccharomyces cerevisiae Proteins
keywords: Bcd1p; Hit1p; NMR structure; box C/D snoRNA; zinc finger protein