title: Functional analysis of the subunits of the chromatin assembly factor RSF.
abstract: The human ISWI-containing factor RSF (for remodeling and spacing factor) is composed of two subunits: the ATPase hSNF2H and p325 (Rsf-1), a protein encoded by a novel human gene. We previously showed that RSF mediates nucleosome deposition and generates regularly spaced nucleosome arrays. Here we report the characterization of the largest subunit of RSF, Rsf-1. We found that Rsf-1 is a highly acidic protein containing a plant homology domain. The present study includes the cloning of Rsf-1, the preparation of recombinant RSF, and the dissection of the role of each subunit in the chromatin assembly reaction. The sequence of the gene for Rsf-1 includes a recently characterized cDNA, HBXAP; postulated to be involved in the transcriptional regulation of the hepatitis B virus. HBXAP actually contains a 252-amino-acid truncation of the amino terminus of Rsf-1. Finally, comparison of HBXAP and Rsf-1 properties shows that they are functionally different.
authors.affiliation: Howard Hughes Medical Institute, University of Medicine and Dentistry of New JerseyRobert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.
authors.forename: Alejandra
authors.initials: A
authors.lastname: Loyola
authors.forename: Jing-Yi
authors.initials: JY
authors.lastname: Huang
authors.forename: Gary
authors.initials: G
authors.lastname: LeRoy
authors.forename: Sherrie
authors.initials: S
authors.lastname: Hu
authors.forename: Yuh-Hwa
authors.initials: YH
authors.lastname: Wang
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Donnelly
authors.forename: William S
authors.initials: WS
authors.lastname: Lane
authors.forename: Sheng-Chung
authors.initials: SC
authors.lastname: Lee
authors.forename: Danny
authors.initials: D
authors.lastname: Reinberg
mesh_terms: D000251:Adenosine Triphosphatases / Q000032:analysis / Q000502:physiology; D000595:Amino Acid Sequence; D002843:Chromatin / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D002868:Chromosomal Proteins, Non-Histone / Q000502:physiology; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism
chemical_list: D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones; D009687:Nuclear Proteins; C452851:RSF1 protein, human; D011994:Recombinant Proteins; D015534:Trans-Activators; D000251:Adenosine Triphosphatases; C409462:SMARCA5 protein, human