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doi:10.1534/genetics.105.047654
A more recent version of this article appeared on February 1, 2006.
REGULAR RESEARCH PAPERS |
Cytogenetic and molecular evaluation of centromere-associated DNA sequences from a marsupial (Macropodidae: Macropus rufogriseus) X chromosome
Kira Bulazel 1, Cushla Metcalfe 2, Gianni Ferreri 1, Jingwei Yu 3, Mark Eldridge 4 and Rachel O'Neill 1*
1 University of Connecticut
2 Macquarie University
3 University of California
4 Macquare University
* To whom correspondence should be addressed. E-mail: rachel.oneill{at}uconn.edu.
Submitted on July 1, 2005
Revised on August 16, 2005
Accepted on 4 October 2005
The constitution of the centromeric portions of the sex chromosomes of the red-necked wallaby, Macropus rufogriseus (family Macropodidae, subfamily Macropodinae), were investigated to develop an overview of the sequence composition of centromeres in a marsupial genome that harbors large amounts of centric and pericentric heterochromatin. The large, C-band positive centromeric region of the X chromosome was microdissected and the isolated DNA was microcloned. Further sequence and cytogenetic analyses of three representative clones show that all chromosomes in this species carry a 178bp satellite sequence containing a CENP-B DNA binding domain (CENP-B box) shown herein to selectively bind marsupial CENP-B protein. Two other repeats isolated in this study localize specifically to the sex chromosomes yet differ in copy number and intrachromosomal distribution. Immunocytohistochemistry assays with anti-CENP-E, anti-CREST, anti-CENP-B, and anti-trimethyl-H3K9 antibodies defined a restricted, point localization of the outer kinetochore at the functional centromere within an enlarged, pericentric and heterochromatic region. The distribution of these repeated sequences within the karyotype of this species, coupled with the apparent high copy number of these sequences, indicates a capacity for retention of large amounts of centromere-associated DNA in the genome of M. rufogriseus.
Key Words: CENP-B, X chromosome, centromere satellite, marsupial, sex chromosomes
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