High-resolution mapping of centromeric protein association using APEX-chromatin fibers.

ABSTRACT:

Background

The centromere is a specialized chromosomal locus that forms the basis for the assembly of a multi-protein complex, the kinetochore and ensures faithful chromosome segregation during every cell division. The repetitive nature of the underlying centromeric sequence represents a major obstacle for high-resolution mapping of protein binding using methods that rely on annotated genomes. Here, we present a novel microscopy-based approach called "APEX-chromatin fibers" for localizing protein binding over the repetitive centromeric sequences at kilobase resolution.

Results

By fusing centromere factors of interest to ascorbate peroxidase, we were able to label their binding profiles on extended chromatin fibers with biotin marks. We applied APEX-chromatin fibers to at lea

SUBMITTER: Kyriacou E 

PROVIDER: S-EPMC6238281 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature