APC/C-mediated degradation of dsRNA-binding protein 4 (DRB4) involved in RNA silencing.

<h4>Background</h4>Selective protein degradation via the ubiquitin-26S proteasome is a major mechanism underlying DNA replication and cell division in all Eukaryotes. In particular, the APC/C (Anaphase Promoting Complex or Cyclosome) is a master ubiquitin protein ligase (E3) that targets...

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Autores principales:	Katia Marrocco, Marie-Claire Criqui, Jérôme Zervudacki, Gregory Schott, Herfried Eisler, Aude Parnet, Patrice Dunoyer, Pascal Genschik
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1b6eb417dd9f4bcbbd7018e5882cae3f
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spelling	oai:doaj.org-article:1b6eb417dd9f4bcbbd7018e5882cae3f2021-11-18T07:21:10ZAPC/C-mediated degradation of dsRNA-binding protein 4 (DRB4) involved in RNA silencing.1932-620310.1371/journal.pone.0035173https://doaj.org/article/1b6eb417dd9f4bcbbd7018e5882cae3f2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22545099/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Selective protein degradation via the ubiquitin-26S proteasome is a major mechanism underlying DNA replication and cell division in all Eukaryotes. In particular, the APC/C (Anaphase Promoting Complex or Cyclosome) is a master ubiquitin protein ligase (E3) that targets regulatory proteins for degradation allowing sister chromatid separation and exit from mitosis. Interestingly, recent work also indicates that the APC/C remains active in differentiated animal and plant cells. However, its role in post-mitotic cells remains elusive and only a few substrates have been characterized.<h4>Methodology/principal findings</h4>In order to identify novel APC/C substrates, we performed a yeast two-hybrid screen using as the bait Arabidopsis APC10/DOC1, one core subunit of the APC/C, which is required for substrate recruitment. This screen identified DRB4, a double-stranded RNA binding protein involved in the biogenesis of different classes of small RNA (sRNA). This protein interaction was further confirmed in vitro and in plant cells. Moreover, APC10 interacts with DRB4 through the second dsRNA binding motif (dsRBD2) of DRB4, which is also required for its homodimerization and binding to its Dicer partner DCL4. We further showed that DRB4 protein accumulates when the proteasome is inactivated and, most importantly, we found that DRB4 stability depends on APC/C activity. Hence, depletion of Arabidopsis APC/C activity by RNAi leads to a strong accumulation of endogenous DRB4, far beyond its normal level of accumulation. However, we could not detect any defects in sRNA production in lines where DRB4 was overexpressed.<h4>Conclusions/significance</h4>Our work identified a first plant substrate of the APC/C, which is not a regulator of the cell cycle. Though we cannot exclude that APC/C-dependent degradation of DRB4 has some regulatory roles under specific growth conditions, our work rather points to a housekeeping function of APC/C in maintaining precise cellular-protein concentrations and homeostasis of DRB4.Katia MarroccoMarie-Claire CriquiJérôme ZervudackiGregory SchottHerfried EislerAude ParnetPatrice DunoyerPascal GenschikPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 4, p e35173 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Katia Marrocco Marie-Claire Criqui Jérôme Zervudacki Gregory Schott Herfried Eisler Aude Parnet Patrice Dunoyer Pascal Genschik APC/C-mediated degradation of dsRNA-binding protein 4 (DRB4) involved in RNA silencing.
description	<h4>Background</h4>Selective protein degradation via the ubiquitin-26S proteasome is a major mechanism underlying DNA replication and cell division in all Eukaryotes. In particular, the APC/C (Anaphase Promoting Complex or Cyclosome) is a master ubiquitin protein ligase (E3) that targets regulatory proteins for degradation allowing sister chromatid separation and exit from mitosis. Interestingly, recent work also indicates that the APC/C remains active in differentiated animal and plant cells. However, its role in post-mitotic cells remains elusive and only a few substrates have been characterized.<h4>Methodology/principal findings</h4>In order to identify novel APC/C substrates, we performed a yeast two-hybrid screen using as the bait Arabidopsis APC10/DOC1, one core subunit of the APC/C, which is required for substrate recruitment. This screen identified DRB4, a double-stranded RNA binding protein involved in the biogenesis of different classes of small RNA (sRNA). This protein interaction was further confirmed in vitro and in plant cells. Moreover, APC10 interacts with DRB4 through the second dsRNA binding motif (dsRBD2) of DRB4, which is also required for its homodimerization and binding to its Dicer partner DCL4. We further showed that DRB4 protein accumulates when the proteasome is inactivated and, most importantly, we found that DRB4 stability depends on APC/C activity. Hence, depletion of Arabidopsis APC/C activity by RNAi leads to a strong accumulation of endogenous DRB4, far beyond its normal level of accumulation. However, we could not detect any defects in sRNA production in lines where DRB4 was overexpressed.<h4>Conclusions/significance</h4>Our work identified a first plant substrate of the APC/C, which is not a regulator of the cell cycle. Though we cannot exclude that APC/C-dependent degradation of DRB4 has some regulatory roles under specific growth conditions, our work rather points to a housekeeping function of APC/C in maintaining precise cellular-protein concentrations and homeostasis of DRB4.
format	article
author	Katia Marrocco Marie-Claire Criqui Jérôme Zervudacki Gregory Schott Herfried Eisler Aude Parnet Patrice Dunoyer Pascal Genschik
author_facet	Katia Marrocco Marie-Claire Criqui Jérôme Zervudacki Gregory Schott Herfried Eisler Aude Parnet Patrice Dunoyer Pascal Genschik
author_sort	Katia Marrocco
title	APC/C-mediated degradation of dsRNA-binding protein 4 (DRB4) involved in RNA silencing.
title_short	APC/C-mediated degradation of dsRNA-binding protein 4 (DRB4) involved in RNA silencing.
title_full	APC/C-mediated degradation of dsRNA-binding protein 4 (DRB4) involved in RNA silencing.
title_fullStr	APC/C-mediated degradation of dsRNA-binding protein 4 (DRB4) involved in RNA silencing.
title_full_unstemmed	APC/C-mediated degradation of dsRNA-binding protein 4 (DRB4) involved in RNA silencing.
title_sort	apc/c-mediated degradation of dsrna-binding protein 4 (drb4) involved in rna silencing.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/1b6eb417dd9f4bcbbd7018e5882cae3f
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_version_	1718423582124539904

APC/C-mediated degradation of dsRNA-binding protein 4 (DRB4) involved in RNA silencing.

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