Replication of ribosomal DNA in Arabidopsis occurs both inside
and outside the nucleolus during S phase progression

J 2018

Replication of ribosomal DNA in Arabidopsis occurs both inside and outside the nucleolus during S phase progression

DVOŘÁČKOVÁ, Martina; B. RAPOSO; Petr MATULA; J. FUCHS; V. SCHUBERT et al.

Basic information

Original name

Replication of ribosomal DNA in Arabidopsis occurs both inside and outside the nucleolus during S phase progression

Authors

DVOŘÁČKOVÁ, Martina ; B. RAPOSO; Petr MATULA; J. FUCHS; V. SCHUBERT; Vratislav PEŠKA; B. DESVOYES; C. GUTIERREZ and Jiří FAJKUS

Edition

Journal of Cell Science, Cambridge, Company of Biologists Ltd. 2018, 0021-9533

Other information

Language

English

Type of outcome

Article in a journal

Field of Study

10601 Cell biology

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

is not subject to a state or trade secret

References:

URL

Impact factor

Impact factor: 4.517

Marked to be transferred to RIV

Yes

RIV identification code

RIV/00216224:14740/18:00100849

Organization unit

Central European Institute of Technology

DOI

https://doi.org/10.1242/jcs.202416

UT WoS

000424842400004

EID Scopus

2-s2.0-85041181109

Keywords in English

A. thaliana rDNA; Replication; Nucleolus; Flow cytometry; Structured illumination microscopy

Abstract

In the original language

Ribosomal RNA genes (rDNA) have been used as valuable experimental systems in numerous studies. Here, we focus on elucidating the spatiotemporal organisation of rDNA replication in Arabidopsis thaliana. To determine the subnuclear distribution of rDNA and the progression of its replication during the S phase, we apply 5-ethynyl-2'-deoxyuridine (EdU) labelling, fluorescence-activated cell sorting, fluorescence in situ hybridization and structured illumination microscopy. We show that rDNA is replicated inside and outside the nucleolus, where active transcription occurs at the same time. Nascent rDNA shows a maximum of nucleolar associations during early S phase. In addition to EdU patterns typical for early or late S phase, we describe two intermediate EdU profiles characteristic for mid S phase. Moreover, the use of lines containing mutations in the chromatin assembly factor-1 gene fas1 and wild-type progeny of fas1xfas2 crosses depleted of inactive copies allows for selective observation of the replication pattern of active rDNA. High-resolution data are presented, revealing the culmination of replication in the mid S phase in the nucleolus and its vicinity. Taken together, our results provide a detailed snapshot of replication of active and inactive rDNA during S phase progression.

Links

GJ16-04166Y, research and development project

Name: Chromatin asociované faktory podílející se na regulaci 45SrDNA v Arabidopsis

Investor: Czech Science Foundation

LH15189, research and development project

Name: Struktura a dynamika 45S rDNA lokusů v Arabidopsis thaliana

Investor: Ministry of Education, Youth and Sports of the CR

LM2015062, research and development project

Name: Národní infrastruktura pro biologické a medicínské zobrazování

Investor: Ministry of Education, Youth and Sports of the CR

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

Cite

DVOŘÁČKOVÁ, Martina; B. RAPOSO; Petr MATULA; J. FUCHS; V. SCHUBERT; Vratislav PEŠKA; B. DESVOYES; C. GUTIERREZ and Jiří FAJKUS. Replication of ribosomal DNA in Arabidopsis occurs both inside and outside the nucleolus during S phase progression. Journal of Cell Science. Cambridge: Company of Biologists Ltd., 2018, vol. 131, No 2, p. nestránkováno, 12 pp. ISSN 0021-9533. Available from: https://doi.org/10.1242/jcs.202416.

@article{1410141,
   author = {Dvořáčková, Martina and Raposo, B. and Matula, Petr and Fuchs, J. and Schubert, V. and Peška, Vratislav and Desvoyes, B. and Gutierrez, C. and Fajkus, Jiří},
   article_location = {Cambridge},
   article_number = {2},
   doi = {https://doi.org/10.1242/jcs.202416},
   keywords = {A. thaliana rDNA; Replication; Nucleolus; Flow cytometry; Structured illumination microscopy},
   language = {eng},
   issn = {0021-9533},
   journal = {Journal of Cell Science},
   title = {Replication of ribosomal DNA in Arabidopsis occurs both inside and outside the nucleolus during S phase progression},
   url = {http://jcs.biologists.org/content/131/2/jcs202416},
   volume = {131},
   year = {2018}
}

TY  - JOUR
ID  - 1410141
AU  - Dvořáčková, Martina - Raposo, B. - Matula, Petr - Fuchs, J. - Schubert, V. - Peška, Vratislav - Desvoyes, B. - Gutierrez, C. - Fajkus, Jiří
PY  - 2018
TI  - Replication of ribosomal DNA in Arabidopsis occurs both inside and outside the nucleolus during S phase progression
JF  - Journal of Cell Science
VL  - 131
IS  - 2
SP  - nestránkováno
EP  - nestránkováno
PB  - Company of Biologists Ltd.
SN  - 00219533
KW  - A. thaliana rDNA
KW  - Replication
KW  - Nucleolus
KW  - Flow cytometry
KW  - Structured illumination microscopy
UR  - http://jcs.biologists.org/content/131/2/jcs202416
L2  - http://jcs.biologists.org/content/131/2/jcs202416
N2  - Ribosomal RNA genes (rDNA) have been used as valuable experimental systems in numerous studies. Here, we focus on elucidating the spatiotemporal organisation of rDNA replication in Arabidopsis thaliana. To determine the subnuclear distribution of rDNA and the progression of its replication during the S phase, we apply 5-ethynyl-2'-deoxyuridine (EdU) labelling, fluorescence-activated cell sorting, fluorescence in situ hybridization and structured illumination microscopy. We show that rDNA is replicated inside and outside the nucleolus, where active transcription occurs at the same time. Nascent rDNA shows a maximum of nucleolar associations during early S phase. In addition to EdU patterns typical for early or late S phase, we describe two intermediate EdU profiles characteristic for mid S phase. Moreover, the use of lines containing mutations in the chromatin assembly factor-1 gene fas1 and wild-type progeny of fas1xfas2 crosses depleted of inactive copies allows for selective observation of the replication pattern of active rDNA. High-resolution data are presented, revealing the culmination of replication in the mid S phase in the nucleolus and its vicinity. Taken together, our results provide a detailed snapshot of replication of active and inactive rDNA during S phase progression.
ER  -

DVOŘÁČKOVÁ, Martina; B. RAPOSO; Petr MATULA; J. FUCHS; V. SCHUBERT; Vratislav PEŠKA; B. DESVOYES; C. GUTIERREZ and Jiří FAJKUS. Replication of ribosomal DNA in Arabidopsis occurs both inside and outside the nucleolus during S phase progression. \textit{Journal of Cell Science}. Cambridge: Company of Biologists Ltd., 2018, vol.~131, No~2, p.~nestránkováno, 12 pp. ISSN~0021-9533. Available from: https://doi.org/10.1242/jcs.202416.

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