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SUV39h-independent association of HP1β with fibrillarin-positive nucleolar regions

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Abstract

Heterochromatin protein 1 (HP1), which binds to sites of histone H3 lysine 9 (H3K9) methylation, is primarily responsible for gene silencing and the formation of heterochromatin. We observed that HP1β is located in both the chromocenters and fibrillarin-positive nucleoli interiors. However, HP1α and HP1γ occupied fibrillarin-positive compartments to a lesser extent, corresponding to the distinct levels of HP1 subtypes at the promoter of rDNA genes. Deficiency of histone methyltransferases SUV39h and/or inhibition of histone deacetylases (HDACi) decreased HP1β and H3K9 trimethylation at chromocenters, but not in fibrillarin-positive regions that co-localized with RNA polymerase I. Similarly, SUV39h- and HDACi-dependent nucleolar rearrangement and inhibition of rDNA transcription did not affect the association between HP1β and fibrillarin. Moreover, the presence of HP1β in nucleoli is likely connected with transcription of ribosomal genes and with the role of fibrillarin in nucleolar processes.

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Abbreviations

μl:

Microliter(s)

μm:

Micrometer(s)

3D:

Three-dimensional

BSA:

Bovine serum albumin

CD:

Chromodomain

CSD:

C-terminal chromoshadow domain

DMEM:

Dulbecco's modified Eagle's medium

dn:

Double null

Dnmt:

DNA methyltransferase

FITC:

Fluorescein isothiocyanate

GFP:

Green fluorescence protein

h:

Hour

H3:

Histone H3

H1, H2A, H2B, H4:

histones H1, H2A, H2B, H4

HDACs:

Histone deacetylases

HMTs:

Histone methyltransferases

HP1:

Heterochromatin protein 1

K (K4, K9, K27...):

Lysine

MEFs:

Mouse embryonic fibroblasts

MeCP2:

Methyl CpG-binding protein

PBS:

Phosphate-buffered saline

PML:

Promyelocytic leukemia bodies

RT:

Room temperature

RNA Pol I:

RNA polymerase I

snoRNAs:

Small nucleolar RNAs

wt:

Wild type

TSA:

Trichostatin A

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Acknowledgments

This work was supported by the research projects: LC535, LC06027, ME919, AVOZ50040702, AVOZ50040507, 2B06052, and 1QS500040508. We thank Prof. Thomas Jenuwein and Dr. Suzane Opravil for providing us the SUV39h-deficient cells, and Dr. Teresa Sullivan and Prof. Collin L. Stewart for LMNA (dn) fibroblasts. We also thank Dr. R. Santoro for providing the rDNA and 28S rDNA primer sequences and Dr. Monika Lachner for the valuable comments of our manuscript. GFP-HP1β plasmid was a generous gift from Dr. Tom Misteli, Laboratory of Receptor Biology and Gene Expression, National Institute of Health, Bethesda. We are also grateful to Dr. Laney Weber from BioScience Writers (Houston, TX, USA) and SPi editing service for the critical revision of our manuscript. Many thanks to Dr. Joost Zomerdijk (from Wellcome Trust Centre for Gene Regulation and Expression, College of Life Sciences, University of Dundee, UK) for recommending the antibody against the large RNA Pol I subunit (RPA194).

Conflict of interest statement

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Institute of Biophysics, Academy of Sciences of the Czech Republic, v.v.i., Královopolská 135, CZ-612 65, Brno, Czech Republic

    Andrea Harničarová Horáková, Eva Bártová, Gabriela Galiová, Radka Uhlířová & Stanislav Kozubek

  2. Faculty of Informatics, Masaryk University, Botanická 68a, CZ-602 00, Brno, Czech Republic

    Pavel Matula

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  1. Andrea Harničarová Horáková
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  2. Eva Bártová
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  5. Pavel Matula
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Correspondence to Eva Bártová.

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Communicated by T. Misteli

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Harničarová Horáková, A., Bártová, E., Galiová, G. et al. SUV39h-independent association of HP1β with fibrillarin-positive nucleolar regions. Chromosoma 119, 227–241 (2010). https://doi.org/10.1007/s00412-009-0252-2

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