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Molecular and Cellular Biology, November 2002, p. 7553-7561, Vol. 22, No. 21
0270-7306/02/$04.00+0 DOI: 10.1128/MCB.22.21.7553-7561.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
Centromere Targeting Element within the Histone Fold Domain of Cid
Danielle Vermaak,1 Hillary S. Hayden,1,
and Steven Henikoff1,2*
Howard Hughes Medical Institute,2
Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, Washington 981091
Received 29 May 2002/
Returned for modification 18 July 2002/
Accepted 30 July 2002
Centromeres require specialized nucleosomes; however, the mechanism of localization is unknown. Drosophila sp. centromeric nucleosomes contain the Cid H3-like protein. We have devised a strategy for identifying elements within Cid responsible for its localization to centromeres. By expressing Cid from divergent Drosophila species fused to green fluorescent protein in Drosophila melanogaster cells, we found that D. bipectinata Cid fails to localize to centromeres. Cid chimeras consisting of the D. bipectinata histone fold domain (HFD) replaced with segments from D. melanogaster identified loop I of the HFD as being critical for targeting to centromeres. Conversely, substitution of D. bipectinata loop I into D. melanogaster abolished centromeric targeting. In either case, loop I was the only segment capable of conferring targeting. Within loop I, we identified residues that are critical for targeting. Most mutations of conserved residues abolished targeting, and length reductions were deleterious. Taken together with the fact that H3 loop I makes numerous contacts with DNA and with the adaptive evolution of Cid, our results point to the importance of DNA specificity for targeting. We suggest that the process of deposition of (Cid.H4)2 tetramers allows for discriminating contacts to be made between loop I and DNA, providing the specificity needed for targeting.
* Corresponding author. Mailing address: Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. N., Seattle, WA 98109. Phone: (206) 667-4515. Fax: (206) 667-5889. E-mail:
steveh{at}fhcrc.org.
Present address: LifeSpan BioSciences, Inc., Seattle, WA 98121.
Molecular and Cellular Biology, November 2002, p. 7553-7561, Vol. 22, No. 21
0022-538X/02/$04.00+0 DOI: 10.1128/MCB.22.21.7553-7561.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
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