A centromeric DNA sequence colocalized with a centromere-specific histone H3 in tobacco.
Centromeres play an important role in segregating chromosomes into daughter cells, and centromeric DNA assembles specific proteins to form a complex referred to as the kinetochore. Among these proteins, centromere-specific histone H3 (CENH3) is one of the most characterized and found to be located only on active centromeres. We isolated four different CENH3-coding complementary DNAs (cDNAs), two from Nicotiana tabaccum and one each from the ancestral diploid species, Nicotiana sylvestris and Nicotiana tomentosiformis and raised an antibody against N-terminal amino acid sequences deduced from the cDNAs. Immunostaining with the antibody revealed the preferential centromere localization, indicating that the cDNAs cloned in this study encode authentic tobacco CENH3. A tobacco centromeric DNA sequence (Nt2-7) was also identified by chromatin immunoprecipitation cloning using the antibody.
Nagaki
Kiyotaka
K
Research Institute for Bioresources, Okayama University, Kurashiki, Japan. nagaki@rib.okayama-u.ac.jp
Kashihara
Kazunari
K
Murata
Minoru
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D002503:Centromere / Q000235:genetics / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D018744:DNA, Plant / Q000235:genetics* / Q000378:metabolism*; D017343:Genes, Plant; D006657:Histones / Q000378:metabolism*; D017404:In Situ Hybridization, Fluorescence; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism*
D018744:DNA, Plant; D006657:Histones; D010940:Plant Proteins