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Cohesin gene defects may impair sister chromatid alignment and genome stability in Arabidopsis thaliana

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Abstract

In contrast to yeast, plant interphase nuclei often display incomplete alignment (cohesion) along sister chromatid arms. Sister chromatid cohesion mediated by the multi-subunit cohesin complex is essential for correct chromosome segregation during nuclear divisions and for DNA recombination repair. The cohesin complex consists of the conserved proteins SMC1, SMC3, SCC3, and an α-kleisin subunit. Viable homozygous mutants could be selected for the Arabidopsis thaliana α-kleisins SYN1, SYN2, and SYN4, which can partially compensate each other. For the kleisin SYN3 and for the single-copy genes SMC1, SMC3, and SCC3, only heterozygous mutants were obtained that displayed between 77% and 97% of the wild-type transcript level. Compared to wild-type nuclei, sister chromatid alignment was significantly decreased along arms in 4C nuclei of the homozygous syn1 and syn4 and even of the heterozygous smc1, smc3, scc3, and syn3 mutants. Knocking out SYN1 and SYN4 additionally impaired sister centromere cohesion. Homozygous mutants of SWITCH1 (required for meiotic sister chromatid alignment) displayed sterility and decreased sister arm alignment. For the cohesin loading complex subunit SCC2, only heterozygous mutants affecting sister centromere alignment were obtained. Defects of the α-kleisin SYN4, which impair sister chromatid alignment in 4C differentiated nuclei, do apparently not disturb alignment during prometaphase nor cause aneuploidy in meristematic cells. The syn2, 3, 4 scc3 and swi1 mutants display a high frequency of anaphases with bridges (~10% to >20% compared to 2.6% in wild type). Our results suggest that (a) already a slight reduction of the average transcript level in heterozygous cohesin mutants may cause perturbation of cohesion, at least in some leaf cells at distinct loci; (b) the decreased sister chromatid alignment in cohesin mutants can obviously not fully be compensated by other cohesion mechanisms such as DNA concatenation; (c) some cohesin genes, in addition to cohesion, might have further essential functions (e.g., for genome stability, apparently by facilitating correct recombination repair of double-strand breaks).

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Acknowledgments

We thank Martina Kühne, Joachim Bruder, and Rita Schubert for technical assistance and Andreas Houben for critical reading the manuscript.

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  1. Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), 06466, Gatersleben, Germany

    Veit Schubert, Andrea Weißleder, Hoda Ali, Jörg Fuchs, Inna Lermontova, Armin Meister & Ingo Schubert

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  1. Veit Schubert
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Correspondence to Veit Schubert.

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Communicated by F. Uhlmann

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Table S1

PCR primers used to identify the T-DNA insertion alleles. (DOC 14 kb)

Table S2

Sequences of the left border junctions of the T-DNA insertion lines. (DOC 13 kb)

Table S3

RT and real-time PCR primers used to amplify transcripts. (DOC 14 kb)

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Schubert, V., Weißleder, A., Ali, H. et al. Cohesin gene defects may impair sister chromatid alignment and genome stability in Arabidopsis thaliana . Chromosoma 118, 591–605 (2009). https://doi.org/10.1007/s00412-009-0220-x

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