Related Experiment Videos
A helix-turn-helix structure unit in human centromere protein B (CENP-B)
J Iwahara1, T Kigawa, K Kitagawa
1Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo 113.
The EMBO Journal
|March 14, 1998
Summary
Centromere Protein B (CENP-B) DNA binding domain RP1 (DBD RP1) forms a helix-turn-helix structure. This structure interacts with essential regions of the CENP-B box DNA, crucial for centromere organization.
Area of Science:
- Molecular Biology
- Chromosomal Biology
- Structural Biology
Background:
- Centromere Protein B (CENP-B) organizes centromeric DNA for kinetochore assembly.
- The N-terminal DNA binding domain (DBD) of CENP-B, comprising RP1 and RP2, binds the CENP-B box sequence.
Purpose of the Study:
- To determine the solution structure of the human CENP-B DBD RP1.
- To investigate the interaction between CENP-B DBD RP1 and the CENP-B box DNA.
Main Methods:
- Multi-dimensional Nuclear Magnetic Resonance (NMR) spectroscopy (1H, 13C, 15N) was used to determine the solution structure.
- NMR chemical shift perturbation was employed to study the protein-DNA interaction.
Main Results:
- The human CENP-B DBD RP1 adopts a helix-turn-helix structure, comprising four helices.
- The overall fold is conserved among eukaryotic DNA binding domains, despite limited sequence homology.
- CENP-B DBD RP1 specifically interacts with key regions of the CENP-B box DNA, including the N-terminal basic region, helix 2, and helix 3.
Conclusions:
- The determined structure provides insights into the molecular basis of CENP-B binding to centromeric DNA.
- Understanding CENP-B DBD RP1 structure and DNA interaction is vital for comprehending centromere formation and chromosomal stability.