Abstract
At the centromere, a network of proteins, the kinetochore, assembles in order to grant correct chromatin segregation. In this study the dynamics and molecular interactions of the inner kinetochore protein CENP-T were analyzed employing a variety of fluorescence microscopy techniques in living human cells. Acceptor-bleaching FRET indicates that CENP-T directly associates with CENP-A and CENP-B. CENP-T exchange into centromeres is restricted to the S-phase of the cell cycle as revealed by FRAP, suggesting a coreplicational loading mechanism, as we have recently also demonstrated for CENP-I. These properties make CENP-T one of the basic inner kinetochore proteins with most further proteins binding downstream, suggesting a fundamental role of CENP-T in kinetochore function.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Autoantigens / metabolism*
-
Base Sequence
-
Cell Cycle
-
Centromere / metabolism*
-
Centromere / ultrastructure
-
Centromere Protein A
-
Centromere Protein B / metabolism*
-
Chromosomal Proteins, Non-Histone / genetics
-
Chromosomal Proteins, Non-Histone / metabolism*
-
DNA Primers / genetics
-
Fluorescence Recovery After Photobleaching
-
Fluorescence Resonance Energy Transfer
-
Green Fluorescent Proteins / genetics
-
Green Fluorescent Proteins / metabolism
-
HeLa Cells
-
Humans
-
Kinetochores / metabolism
-
Kinetochores / ultrastructure
-
Microscopy, Fluorescence
-
Optical Phenomena
-
Recombinant Fusion Proteins / genetics
-
Recombinant Fusion Proteins / metabolism
-
Spectrometry, Fluorescence
Substances
-
Autoantigens
-
CENPA protein, human
-
CENPB protein, human
-
CENPT protein, human
-
Centromere Protein A
-
Centromere Protein B
-
Chromosomal Proteins, Non-Histone
-
DNA Primers
-
Recombinant Fusion Proteins
-
enhanced green fluorescent protein
-
Green Fluorescent Proteins