Limits...
Involvement of the Cohesin Cofactor PDS5 (SPO76) During Meiosis and DNA Repair in Arabidopsis thaliana.

Pradillo M, Knoll A, Oliver C, Varas J, Corredor E, Puchta H, Santos JL - Front Plant Sci (2015)

Bottom Line: Cohesin cofactors contribute to cohesin dynamics and interact with cohesin complexes during cell cycle.In Arabidopsis, AtWAPL proteins are essential during meiosis, however, the role of AtPDS5 remains to be ascertained.Furthermore, this cohesin cofactor could be important for the function of the AtSMC5/AtSMC6 complex.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Genética, Facultad de Biología, Universidad Complutense Madrid, Spain.

ABSTRACT
Maintenance and precise regulation of sister chromatid cohesion is essential for faithful chromosome segregation during mitosis and meiosis. Cohesin cofactors contribute to cohesin dynamics and interact with cohesin complexes during cell cycle. One of these, PDS5, also known as SPO76, is essential during mitosis and meiosis in several organisms and also plays a role in DNA repair. In yeast, the complex Wapl-Pds5 controls cohesion maintenance and colocalizes with cohesin complexes into chromosomes. In Arabidopsis, AtWAPL proteins are essential during meiosis, however, the role of AtPDS5 remains to be ascertained. Here we have isolated mutants for each of the five AtPDS5 genes (A-E) and obtained, after different crosses between them, double, triple, and even quadruple mutants (Atpds5a Atpds5b Atpds5c Atpds5e). Depletion of AtPDS5 proteins has a weak impact on meiosis, but leads to severe effects on development, fertility, somatic homologous recombination (HR) and DNA repair. Furthermore, this cohesin cofactor could be important for the function of the AtSMC5/AtSMC6 complex. Contrarily to its function in other species, our results suggest that AtPDS5 is dispensable during the meiotic division of Arabidopsis, although it plays an important role in DNA repair by HR.

No MeSH data available.


Related in: MedlinePlus

AtPDS5 genes are involved in DNA repair, they are overexpressed after γ-irradiation and their loss of function generates down-regulation of AtSMC6 genes. (A) Expression analysis of AtPDS5 genes after γ-irradiation in WT leaf samples. (B) Expression analysis of AtSMC6A and AtSMC6B in double, triple, and quadruple mutant leaf samples. Transcript levels are relative to non-irradiated WT (discontinuous line) (see Materials and Methods for more details). RQ: relative quantity. ab: Atpds5a Atpds5b; abc: Atpds5a Atpds5b Atpds5c; abce: Atpds5a Atpds5b Atpds5c Atpds5e.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4664637&req=5

Figure 5: AtPDS5 genes are involved in DNA repair, they are overexpressed after γ-irradiation and their loss of function generates down-regulation of AtSMC6 genes. (A) Expression analysis of AtPDS5 genes after γ-irradiation in WT leaf samples. (B) Expression analysis of AtSMC6A and AtSMC6B in double, triple, and quadruple mutant leaf samples. Transcript levels are relative to non-irradiated WT (discontinuous line) (see Materials and Methods for more details). RQ: relative quantity. ab: Atpds5a Atpds5b; abc: Atpds5a Atpds5b Atpds5c; abce: Atpds5a Atpds5b Atpds5c Atpds5e.

Mentions: Exposure of plants to DSB-inducing agents increases transcript levels of genes involved in DNA repair (Culligan et al., 2006). For this reason, we tested by qPCR the expression levels of the different AtPDS5 genes after γ-irradiation treatment. The results obtained revealed increased levels in the expression of these genes at 300 and 500 Gy in leaf tissue (Figure 5A). The most noteworthy increase was observed in AtPDS5E at 500 Gy, the expression of this gene was 10-fold higher than in the untreated control. However, no significant change was observed for the expression of AtPDS5 genes (except a slight increase for AtPDS5E) using samples from buds, containing meiocytes (Supplementary Figure S8A). We also investigated whether the loss of AtPDS5 genes alters the expression of genes required for DNA repair, which includes the kinases AtATM and AtATR, AtRAD50, the ubiquitin ligase AtBRCA1, the recombinase AtRAD51 and the SMC genes AtSMC6A and AtSMC6B (Gallego et al., 2001; Daoudal-Cotterell et al., 2002; Lafarge and Montané, 2003; Li et al., 2004; Culligan and Britt, 2008; Watanabe et al., 2009; Amiard et al., 2010; Pradillo et al., 2012). Small and unremarkable changes (down and up) were observed for AtATR, AtRAD50, AtBRCA1, and AtRAD51 expression in leaf samples. AtATM exhibited a reduced expression in both the triple and the quadruple mutants and all these genes were down-regulated in bud samples (Supplementary Figure S9). However, the most significant result was observed for AtSMC6A and AtSMC6B since the transcripts of these genes in the triple and quadruple mutants were halved and reduced by more than half, respectively (Figure 5B). This reduction was also observed in bud samples (Supplementary Figure S8B).


Involvement of the Cohesin Cofactor PDS5 (SPO76) During Meiosis and DNA Repair in Arabidopsis thaliana.

Pradillo M, Knoll A, Oliver C, Varas J, Corredor E, Puchta H, Santos JL - Front Plant Sci (2015)

AtPDS5 genes are involved in DNA repair, they are overexpressed after γ-irradiation and their loss of function generates down-regulation of AtSMC6 genes. (A) Expression analysis of AtPDS5 genes after γ-irradiation in WT leaf samples. (B) Expression analysis of AtSMC6A and AtSMC6B in double, triple, and quadruple mutant leaf samples. Transcript levels are relative to non-irradiated WT (discontinuous line) (see Materials and Methods for more details). RQ: relative quantity. ab: Atpds5a Atpds5b; abc: Atpds5a Atpds5b Atpds5c; abce: Atpds5a Atpds5b Atpds5c Atpds5e.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664637&req=5

Figure 5: AtPDS5 genes are involved in DNA repair, they are overexpressed after γ-irradiation and their loss of function generates down-regulation of AtSMC6 genes. (A) Expression analysis of AtPDS5 genes after γ-irradiation in WT leaf samples. (B) Expression analysis of AtSMC6A and AtSMC6B in double, triple, and quadruple mutant leaf samples. Transcript levels are relative to non-irradiated WT (discontinuous line) (see Materials and Methods for more details). RQ: relative quantity. ab: Atpds5a Atpds5b; abc: Atpds5a Atpds5b Atpds5c; abce: Atpds5a Atpds5b Atpds5c Atpds5e.
Mentions: Exposure of plants to DSB-inducing agents increases transcript levels of genes involved in DNA repair (Culligan et al., 2006). For this reason, we tested by qPCR the expression levels of the different AtPDS5 genes after γ-irradiation treatment. The results obtained revealed increased levels in the expression of these genes at 300 and 500 Gy in leaf tissue (Figure 5A). The most noteworthy increase was observed in AtPDS5E at 500 Gy, the expression of this gene was 10-fold higher than in the untreated control. However, no significant change was observed for the expression of AtPDS5 genes (except a slight increase for AtPDS5E) using samples from buds, containing meiocytes (Supplementary Figure S8A). We also investigated whether the loss of AtPDS5 genes alters the expression of genes required for DNA repair, which includes the kinases AtATM and AtATR, AtRAD50, the ubiquitin ligase AtBRCA1, the recombinase AtRAD51 and the SMC genes AtSMC6A and AtSMC6B (Gallego et al., 2001; Daoudal-Cotterell et al., 2002; Lafarge and Montané, 2003; Li et al., 2004; Culligan and Britt, 2008; Watanabe et al., 2009; Amiard et al., 2010; Pradillo et al., 2012). Small and unremarkable changes (down and up) were observed for AtATR, AtRAD50, AtBRCA1, and AtRAD51 expression in leaf samples. AtATM exhibited a reduced expression in both the triple and the quadruple mutants and all these genes were down-regulated in bud samples (Supplementary Figure S9). However, the most significant result was observed for AtSMC6A and AtSMC6B since the transcripts of these genes in the triple and quadruple mutants were halved and reduced by more than half, respectively (Figure 5B). This reduction was also observed in bud samples (Supplementary Figure S8B).

Bottom Line: Cohesin cofactors contribute to cohesin dynamics and interact with cohesin complexes during cell cycle.In Arabidopsis, AtWAPL proteins are essential during meiosis, however, the role of AtPDS5 remains to be ascertained.Furthermore, this cohesin cofactor could be important for the function of the AtSMC5/AtSMC6 complex.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Genética, Facultad de Biología, Universidad Complutense Madrid, Spain.

ABSTRACT
Maintenance and precise regulation of sister chromatid cohesion is essential for faithful chromosome segregation during mitosis and meiosis. Cohesin cofactors contribute to cohesin dynamics and interact with cohesin complexes during cell cycle. One of these, PDS5, also known as SPO76, is essential during mitosis and meiosis in several organisms and also plays a role in DNA repair. In yeast, the complex Wapl-Pds5 controls cohesion maintenance and colocalizes with cohesin complexes into chromosomes. In Arabidopsis, AtWAPL proteins are essential during meiosis, however, the role of AtPDS5 remains to be ascertained. Here we have isolated mutants for each of the five AtPDS5 genes (A-E) and obtained, after different crosses between them, double, triple, and even quadruple mutants (Atpds5a Atpds5b Atpds5c Atpds5e). Depletion of AtPDS5 proteins has a weak impact on meiosis, but leads to severe effects on development, fertility, somatic homologous recombination (HR) and DNA repair. Furthermore, this cohesin cofactor could be important for the function of the AtSMC5/AtSMC6 complex. Contrarily to its function in other species, our results suggest that AtPDS5 is dispensable during the meiotic division of Arabidopsis, although it plays an important role in DNA repair by HR.

No MeSH data available.


Related in: MedlinePlus