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Effects of scaffold/matrix alteration on centromeric function and gene expression

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Version 2 2024-06-04, 13:33
Version 1 2019-03-01, 11:24
journal contribution
posted on 2024-06-04, 13:33 authored by Huseyin Sumer, Richard Saffery, Nicholas Wong, Jeffrey CraigJeffrey Craig, KH Andy Choo
We have previously described a 3.5-Mb domain of enhance scaffold/matrix attachment region (S/MAR) at a human neocentromere, and normal expression of underlying genes within this region. We also reported that partial inhibition of histone deacetylation using 33 nmtrichostatin A (TSA) resulted in a shift in the position of the CENP-A-binding domain within the neocentromere, with no noticeable effects on mitotic segregation function. In this study, 33 nM TSA caused a reduction in the size of the enhanced S/MAR domain of one-half to 1.7 Mb. Treatment with a DNA-intercalating drug distamycin A (DST) at 75 microg/ml resulted in a size reduction of the enhanced S/MAR domain at the neocentromere of two-thirds to 1.2 Mb, and that of the CENP-A-binding domain of 40%, from 330 to 196 kb, with no significant shift in the position of the latter domain. Other DST effects include mitotic chromosomal missegregation, reduction in the levels of Topo IIalpha, CENP-A, CENP-C, and HP1alpha, and an increase in mitotic checkpoint protein BubR1. TSA or DST treatment similarly resulted in a significant reduction, by approximately 20 and 50%, respectively, in the size of the enhanced S/MAR domain at the alpha-satellite DNA of a native chromosome 10 centromere. Transcriptional competence within the neocentromere is overall not noticeably altered by either TSA or DST treatment, as is evident from the absence of any significant increase or decrease in the expression levels of 47 underlying genes tested. These results suggest that a substantial contraction of the S/MAR domain may not be deleterious to centromere function, that disruption of the S/MAR domain directly affects the binding properties of a host of scaffold/matrix and centromeric/pericentric proteins, and that the overall competence and regulation of transcription at the neocentromeric chromatin is similar to those found at the corresponding normal genomic sites.

History

Journal

Journal of biological chemistry

Volume

279

Pagination

37631-37639

Location

Rockville, Md.

Open access

  • Yes

ISSN

0021-9258

Language

eng

Publication classification

C1.1 Refereed article in a scholarly journal

Copyright notice

2004, The American Society for Biochemistry and Molecular Biology, Inc.

Issue

36

Publisher

American Society for Biochemistry and Molecular Biology