Mutational analysis of the central centromere targeting domain of human centromere protein C, (CENP-C)

Experimental Cell Research
Kang SongJ Tomkiel

Abstract

Human centromere protein C (CENP-C) is an essential component of the inner kinetochore plate. A central region of CENP-C can bind DNA in vitro and is sufficient for targeting the protein to centromeres in vivo, raising the possibility that this domain mediates centromere localization via direct DNA binding. We performed a detailed molecular dissection of this domain to understand the mechanism by which CENP-C assembles at centromeres. By a combination of PCR mutagenesis and transient expression of GFP-tagged proteins in HeLa cells, we identified mutations that disrupt centromere localization of CENP-C in vivo. These cluster in a 12 amino acid region adjacent to the core domain required for in vitro DNA binding. This region is conserved between human and mouse, but is divergent or absent in invertebrate and plant CENP-C homologues. We suggest that these 12 amino acids are essential to confer specificity to DNA binding by CENP-C in vivo, or to mediate interaction with another as yet unidentified centromere component. A differential yeast two-hybrid screen failed to identify interactions specific to this sequence, but nonetheless identified 14 candidate proteins that interact with the central region of CENP-C. This collection of m...Continue Reading

Citations

Jul 4, 2012·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·Kristina M SmithMichael Freitag
Jul 31, 2009·Molecular Biology of the Cell·Kirstin J MilksAaron F Straight
Feb 10, 2010·Molecular Biology and Evolution·Mary G SchuelerEric D Green
Sep 6, 2005·Genes & Development·Sebastian HeegerChristian F Lehner
Jun 8, 2006·Annual Review of Genomics and Human Genetics·Mary G Schueler, Beth A Sullivan
Sep 4, 2004·Journal of Biology·Paul B TalbertSteven Henikoff
Jul 1, 2015·The Journal of Cell Biology·Kerstin KlareAndrea Musacchio
Jun 23, 2010·The Journal of Cell Biology·Christopher W CarrollAaron F Straight
Jan 27, 2017·Biology·Andrea Musacchio, Arshad Desai
Nov 13, 2003·The Journal of Biological Chemistry·Nobutaka SuzukiHiroshi Masumoto
Sep 3, 2019·EMBO Reports·Ahmad Ali-AhmadNikolina Sekulić
Sep 10, 2013·Cell Cycle·Hidenori KatoYawen Bai

❮ Previous
Next ❯

Related Concepts

Related Feeds

Autoimmune Diseases

Autoimmune diseases occur as a result of an attack by the immune system on the body’s own tissues resulting in damage and dysfunction. There are different types of autoimmune diseases, in which there is a complex and unknown interaction between genetics and the environment. Discover the latest research on autoimmune diseases here.