Functional analysis of the subunits of the chromatin assembly factor RSF.
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.
Loyola
Alejandra
A
Howard Hughes Medical Institute, University of Medicine and Dentistry of New JerseyRobert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.
Huang
Jing-Yi
JY
LeRoy
Gary
G
Hu
Sherrie
S
Wang
Yuh-Hwa
YH
Donnelly
Robert J
RJ
Lane
William S
WS
Lee
Sheng-Chung
SC
Reinberg
Danny
D
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
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