Mechanisms of nuclear lamina growth in interphase

Zhironkina, Oxana A.; Kurchashova, Svetlana Yu; Pozharskaia, Vasilisa A.; Cherepanynets, Varvara D.; Strelkova, Olga S.; Hozak, Pavel; Kireev, Igor I.

doi:10.1007/s00418-016-1419-6

Original Paper
Published: 16 February 2016

Mechanisms of nuclear lamina growth in interphase

Oxana A. Zhironkina¹,
Svetlana Yu Kurchashova¹,
Vasilisa A. Pozharskaia³,
Varvara D. Cherepanynets¹,
Olga S. Strelkova¹,
Pavel Hozak⁴ &
Igor I. Kireev^1,2

Histochemistry and Cell Biology volume 145, pages 419–432 (2016)Cite this article

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Abstract

The nuclear lamina represents a multifunctional platform involved in such diverse yet interconnected processes as spatial organization of the genome, maintenance of mechanical stability of the nucleus, regulation of transcription and replication. Most of lamina activities are exerted through tethering of lamina-associated chromatin domains (LADs) to the nuclear periphery. Yet, the lamina is a dynamic structure demonstrating considerable expansion during the cell cycle to accommodate increased number of LADs formed during DNA replication. We analyzed dynamics of nuclear growth during interphase and changes in lamina structure as a function of cell cycle progression. The nuclear lamina demonstrates steady growth from G1 till G2, while quantitative analysis of lamina meshwork by super-resolution microscopy revealed that microdomain organization of the lamina is maintained, with lamin A and lamin B microdomain periodicity and interdomain gap sizes unchanged. FRAP analysis, in contrast, demonstrated differences in lamin A and B1 exchange rates; the latter showing higher recovery rate in S-phase cells. In order to further analyze the mechanism of lamina growth in interphase, we generated a lamina-free nuclear envelope in living interphase cells by reversible hypotonic shock. The nuclear envelope in nuclear buds formed after such a treatment initially lacked lamins, and analysis of lamina formation revealed striking difference in lamin A and B1 assembly: lamin A reassembled within 30 min post-treatment, whereas lamin B1 did not incorporate into the newly formed lamina at all. We suggest that in somatic cells lamin B1 meshwork growth is coordinated with replication of LADs, and lamin A meshwork assembly seems to be chromatin-independent process.

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Acknowledgments

The authors thank J. Ellenberg and M. Cardoso, who kindly provided plasmids for this work. This research was partially supported by the Institute of Molecular Genetics, Academy of Sciences of the Czech Republic (RVO: 68378050 to PH), the Grant agency of the Czech Republic (16-03403S to PH), Russian Fund for Basic Research (Grants 13-04-00885 and 15-54-78077 to IIK) and Moscow State university Development Program (PNR 5.13).

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Department of Electron Microscopy, A. N. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 1-40 Leninskie Gory, Moscow, Russia
Oxana A. Zhironkina, Svetlana Yu Kurchashova, Varvara D. Cherepanynets, Olga S. Strelkova & Igor I. Kireev
Faculty of Biology, Lomonosov Moscow State University, 1-40 Leninskie Gory, Moscow, Russia
Igor I. Kireev
Faculty of Chemistry, Lomonosov Moscow State University, 1-3 Leninskie Gory, Moscow, Russia
Vasilisa A. Pozharskaia
Institute of Molecular Genetics of the ASCR, v. v. i. Vídeňská 1083, 142 20, Prague 4, Czech Republic
Pavel Hozak

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Oxana A. Zhironkina
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Svetlana Yu Kurchashova
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Vasilisa A. Pozharskaia
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Varvara D. Cherepanynets
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Olga S. Strelkova
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Pavel Hozak
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Igor I. Kireev
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Correspondence to Igor I. Kireev.

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Zhironkina, O.A., Kurchashova, S.Y., Pozharskaia, V.A. et al. Mechanisms of nuclear lamina growth in interphase. Histochem Cell Biol 145, 419–432 (2016). https://doi.org/10.1007/s00418-016-1419-6

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Accepted: 01 February 2016
Published: 16 February 2016
Issue Date: April 2016
DOI: https://doi.org/10.1007/s00418-016-1419-6

Keywords

Nuclear lamina
Microdomains
Interphase
Nucleus
DNA replication
Cell cycle