Centromere-Encoded RNAs Are Integral Components of the Maize Kinetochore

RNA is involved in a variety of chromatin modification events, ranging from large-scale structural rearrangements to subtle local affects. Here, we extend the evidence for RNA-chromatin interactions to the centromere core. The data indicate that maize centromeric retrotransposons (CRMs) and satellit...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 101; no. 45; pp. 15986 - 15991
Main Authors Topp, Christopher N., Zhong, Cathy X., Dawe, R. Kelly, Wessler, Susan R.
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 09.11.2004
National Acad Sciences
Subjects
Base Sequence
Biological Sciences
Centromere - genetics
Centromere - metabolism
Centromeres
Chromatin
Chromosomes
Corn
DNA
DNA, Plant - genetics
Genetics
Histones
Histones - metabolism
Kinetochores
Kinetochores - metabolism
Plant Proteins - metabolism
Retroelements - genetics
Retrotransposons
Ribonucleic acid
RNA
RNA, Plant - genetics
Small interfering RNA
Zea mays
Zea mays - genetics
Zea mays - metabolism
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Summary:RNA is involved in a variety of chromatin modification events, ranging from large-scale structural rearrangements to subtle local affects. Here, we extend the evidence for RNA-chromatin interactions to the centromere core. The data indicate that maize centromeric retrotransposons (CRMs) and satellite repeats (CentC) are not only transcribed, but that nearly half of the CRM and CentC RNA is tightly bound to centromeric histone H3 (CENH3), a key inner kinetochore protein. RNAs from another tandem repeat (180-bp knob sequence) or an abundant euchromatic retroelement (Opie) are undetectable within the same anti-CENH3 immune complexes. Both sense and antisense strands of CRM and CentC, but not small interfering RNAs homologous to either repeat, were found to coimmunoprecipitate with CENH3. The bulk of the immunoprecipitated RNA ranged in size from 40 to 200 nt. These data provide evidence for a pool of protected, single-stranded centromeric RNA within the centromere/kinetochore complex.
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To whom correspondence should be addressed. E-mail: kelly@plantbio.uga.edu.
Author contributions: R.K.D. designed research; C.N.T. and C.X.Z. performed research; R.K.D. and C.N.T. analyzed data; and R.K.D. and C.N.T. wrote the paper.
Communicated by Susan R. Wessler, University of Georgia, Athens, GA, September 28, 2004
Abbreviations: CENH3, centromeric histone H3; RNAi, RNA interference; siRNA, small interfering RNA; ChIP, chromatin immunoprecipitation; MNase, micrococcal nuclease; CR, centromeric retrotransposable; CRM, maize CR; s/n, signal to noise; %IP, percentage of immunoprecipitation.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0407154101