Calreticulin as a new histone binding protein in mitotic chromosomes.
Calreticulin (CRT) is a multifunctional Ca(2+)-binding protein that mainly functions in the endoplasmic reticulum as a molecular chaperone for newly synthesized proteins. Recently we reported the protein composition of human metaphase chromosomes (Uchiyama et al., 2004), which included CRT. Here we describe new characteristics of CRT in vitro as well as its localization on the surface of metaphase chromosomes in vivo. CRT was detected in the chromosomal fraction by Western blotting and its binding partners were identified as core and linker histones by ligand overlay assay. Surface plasmon resonance sensor analyses revealed that CRT is bound to chromatin fibers. Moreover, we found that CRT has both supercoiling activity, which assists core histone assembly into chromatin fibers, and binding ability to histone H2A/H2B dimers and histone H3/H4 tetramers. Unlike the chromosome scaffold proteins, indirect immunofluorescent staining revealed that CRT is located on the surface of metaphase chromosomes. These results suggest that CRT plays a role which involves chromatin dynamics on the surface of mitotic chromosomes.
Kobayashi
S
S
Department of Biotechnology, Graduate School of Engineering, Osaka University, Yamadaoka, Suita, Japan.
Uchiyama
S
S
Sone
T
T
Noda
M
M
Lin
L
L
Mizuno
H
H
Matsunaga
S
S
Fukui
K
K
D037282:Calreticulin / Q000378:metabolism* / Q000502:physiology; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D002875:Chromosomes / Q000378:metabolism*; D019281:Dimerization; D006657:Histones / Q000378:metabolism; D006801:Humans; D008677:Metaphase; D008938:Mitosis; D011485:Protein Binding; D011994:Recombinant Proteins
D037282:Calreticulin; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones; D011994:Recombinant Proteins