The rapidly evolving centromere-specific histone has stringent functional requirements in Arabidopsis thaliana.
Centromeres control chromosome inheritance in eukaryotes, yet their DNA structure and primary sequence are hypervariable. Most animals and plants have megabases of tandem repeats at their centromeres, unlike yeast with unique centromere sequences. Centromere function requires the centromere-specific histone CENH3 (CENP-A in human), which replaces histone H3 in centromeric nucleosomes. CENH3 evolves rapidly, particularly in its N-terminal tail domain. A portion of the CENH3 histone-fold domain, the CENP-A targeting domain (CATD), has been previously shown to confer kinetochore localization and centromere function when swapped into human H3. Furthermore, CENP-A in human cells can be functionally replaced by CENH3 from distantly related organisms including Saccharomyces cerevisiae. We have used cenh3-1 (a null mutant in Arabidopsis thaliana) to replace endogenous CENH3 with GFP-tagged variants. A H3.3 tail domain-CENH3 histone-fold domain chimera rescued viability of cenh3-1, but CENH3's lacking a tail domain were nonfunctional. In contrast to human results, H3 containing the A. thaliana CATD cannot complement cenh3-1. GFP-CENH3 from the sister species A. arenosa functionally replaces A. thaliana CENH3. GFP-CENH3 from the close relative Brassica rapa was targeted to centromeres, but did not complement cenh3-1, indicating that kinetochore localization and centromere function can be uncoupled. We conclude that CENH3 function in A. thaliana, an organism with large tandem repeat centromeres, has stringent requirements for functional complementation in mitosis.
Ravi
Maruthachalam
M
Department of Chemistry and Biochemistry, Universityof California, Los Angeles, CA 90095, USA.
Kwong
Pak N
PN
Menorca
Ron M G
RM
Valencia
Joel T
JT
Ramahi
Joseph S
JS
Stewart
Jodi L
JL
Tran
Robert K
RK
Sundaresan
Venkatesan
V
Comai
Luca
L
Chan
Simon W-L
SW
D017360:Arabidopsis / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000378:metabolism; D002503:Centromere / Q000378:metabolism*; D019143:Evolution, Molecular; D005816:Genetic Complementation Test; D006657:Histones / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D018386:Kinetochores / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013045:Species Specificity; D019076:Transgenes
D029681:Arabidopsis Proteins; D018797:Cell Cycle Proteins; D006657:Histones; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins