AtCTF7 is required for establishment of sister chromatid cohesion and association of cohesin with chromatin during meiosis in Arabidopsis
© Singh et al.; licensee BioMed Central Ltd. 2013
Received: 8 April 2013
Accepted: 5 August 2013
Published: 14 August 2013
The establishment of sister chromatid cohesion followed by its controlled release at the metaphase to anaphase transition is necessary for faithful segregation of chromosomes in mitosis and meiosis. Cohesion is established by the action of Ctf7/Eco1 on the cohesin complex during DNA replication following loading of cohesin onto chromatin by the Scc2-Scc4 complex. Ctf7 is also required for sister chromatid cohesion during repair of DNA double strand breaks. Ctf7 contains an acetyltransferase domain and a zinc finger motif and acetylates conserved lysine residues in the Smc3 subunit of cohesin. In Arabidopsis CTF7 is encoded by a single gene and mutations in AtCTF7 cause embryo lethality indicating that the gene is essential.
To study the function of Ctf7 in plants and to determine its role in sister chromatid cohesion, we constructed a conditional allele of AtCTF7 in Arabidopsis using an inducible RNA interference (RNAi) strategy, so as to avoid the embryo lethality caused by mutations in AtCTF7. We found that induction of RNAi against AtCTF7 caused severe inhibition and defects in growth during vegetative and reproductive stages as well as sterility. AtCTF7-RNAi plants displayed chromosome fragmentation and loss of sister chromatid cohesion during meiosis. Immunostaining for the cohesion subunit AtSCC3 showed a marked reduction in association of cohesin with chromatin during meiosis in AtCTF7-RNAi plants.
We find that AtCTF7 is essential for sister chromatid cohesion during meiosis in Arabidopsis and is required for association of cohesin with chromatin in prophase of meiosis.
KeywordsPlant meiosis Chromosome organization Synapsis Conditional RNAi Gametogenesis DNA repair
Proper chromosome segregation during cell division requires that sister chromatids produced by DNA replication are held together until their controlled separation at anaphase. This function is accomplished by the cohesin complex, whose conserved core subunits consist of the Structural Maintenance of Chromosome (SMC) proteins Smc1 and Smc3, the Sister Chromatid Cohesion (SCC) protein Scc3, and the α-kleisin protein Scc1 . According to the ring model of cohesin action, Smc1 and Smc3 interact to form a V shaped heterodimer, closed by Scc1 with the help of Scc3, to form a ring that is considered to entrap sister chromatids and hold them physically together [2, 3]. Cohesion is released at anaphase by the cleavage of Scc1 by separase, a protease that is activated by the anaphase promoting complex/cyclosome (APC/C) .
In Saccharomyces cerevisiae, cohesion is established by Ctf7/Eco1, after cohesin has been loaded on chromatin by the Scc2-Scc4 complex [4–6]. Ctf7 establishes cohesion during S phase, and interacts with components of the DNA replication machinery, including PCNA and RFC [5–7]. These results led to a model in which sister chromatid cohesion is established concomitantly with DNA replication . Ctf7 encodes a zinc finger protein with an active acetyltransferase domain, and it was found that Ctf7 acetylation of Smc3 on conserved lysines, was critical for establishment of cohesion by counteracting the Wpl1-Pds5 complex in preventing establishment of cohesion [9–14]. Establishment of cohesion has been suggested to occur in concert with lagging strand synthesis , and Smc3 acetylation leading to establishment of functional cohesion occurs only in association with replication . Recycling of the Smc3 subunit is aided by deacetylation by Hos1 following cleavage of Scc1 by separase to release cohesion at the metaphase to anaphase transition, and is important for establishment of cohesion [17–19]. The eso1-H17 mutant in Schizosaccharomyces pombe exhibits delayed mitosis as a result of activation of the spindle checkpoint, and defective segregation of chromosomes in mitosis . In Drosophila, mutations in Deco result in altered distribution of cohesin at metaphase, and premature entry into anaphase . In humans, mutations in ESCO2 cause Roberts syndrome which results from a deficiency of cohesion around the centromeres, and encompasses a number of developmental abnormalities as well as mental retardation and renal and cardiac dysfunction .
The machinery for establishment of cohesion is conserved in Arabidopsis, and homologues of Scc2 and Ctf7 have been identified and functionally characterized. Mutations in AtSCC2 and AtCTF7 result in embryo lethality, however AtCTF7 is dispensable for endosperm growth [23, 24]. Interestingly, AtCTF7 was found to possess acetyltransferase activity in vitro, and could complement the yeast ctf7-203 mutant, suggesting conserved biochemical function with its yeast counterpart . By using a conditional RNA interference (RNAi) approach, it has been demonstrated that AtSCC2 is required during meiosis for sister chromatid cohesion, chromosomal axis formation and synapsis between homologues . The function of AtCTF7 in establishment of sister chromatid cohesion in planta remains to be shown.
Here, we used a conditional RNAi approach to examine the role of AtCTF7 in sister chromatid cohesion, and to analyze the effects of the loss of AtCTF7 during vegetative and reproductive development. We found that downregulation of AtCTF7 severely inhibited growth during vegetative and reproductive stages, and resulted in both male and female sterility. During meiosis, AtCTF7-RNAi lines displayed typical loss of cohesion phenotypes, including abnormal chromosome organization, impaired chromosome synapsis and DNA fragmentation. Consistent with an expected involvement of AtCTF7 in cohesion, we found that sister chromatid cohesion was lost at both chromosome arms and centromeres in AtCTF7-RNAi plants. Finally, we found that AtSCC3 localization on chromatin was compromised during meiosis in AtCTF7-RNAi plants, indicating that AtCTF7 is required for association of cohesin with chromosomes in Arabidopsis, a feature that appears to be similar to Drosophila, where Deco is required for Scc1 association with chromosomes during M phase . Overall, our results establish an essential role for AtCTF7 in vegetative development and in sister chromatid cohesion during meiosis.
AtCTF7 is required for growth during vegetative and reproductive stages
These results indicate that AtCTF7 is expressed in dividing cells and is required for normal development and growth during both vegetative and reproductive stages.
Knockdown of AtCTF7 results in defects in sister chromatid cohesion and chromosome organization during meiosis
Reduced association of cohesin with chromatin in AtCTF7-RNAi plants
Ctf7/Eco1 proteins have been shown to control establishment of cohesion in yeast, Drosophila, and mammals [5, 6, 20, 21, 29]. In the case of plants, the Arabidopsis homolog of Ctf7 (AtCTF7) has been shown to be required for embryo development but not required for development of the endosperm , leaving open the possibility of a Ctf7-independent mechanism for sister chromatid segregation operating in meiosis. Evidence for a Ctf7-independent mechanism for sister chromatid segregation in yeast is based on the viability of an eco1∆ wpl1∆strain . A dosage dependent role for Ctf7 in meiosis in yeast has been suggested based on haplo-insufficiency during sporulation . Establishment of sister chromatid cohesion in meiosis in yeast may therefore be more sensitive to reduced dosage of Ctf7 than in mitosis. Alternatively, acetylation of other proteins by Ctf7 during meiosis may also be involved . A role for Ctf7 in meiosis is also suggested from an examination of the localization and regulation of murine ESCO2 , however a requirement for Ctf7 in sister chromatid cohesion specifically during meiosis remains to be established. In this study we have shown using Arabidopsis as a model, that AtCTF7 is also required for sister chromatid cohesion in meiosis.
The conditional RNAi approach to examine the function of AtCTF7 in plants revealed defects in sister chromatid cohesion in meiosis. The establishment of sister chromatid cohesion in both arm and centromeric regions during meiosis was dependent upon AtCTF7. The meiotic phenotypes comprised defects early in prophase which presented as discontinuities in the thread-like appearance characteristic of leptotene and zygotene stages. In the most severe cases, the thread-like structure was largely absent and the chromatin appeared highly fragmented. At later prophase stages, the fragmented phenotype was further apparent by the presence of a large number of separated and condensed chromatin fragments. The results are consistent with a failure (in meiosis) to repair double strand breaks for which Ctf7 is known to be required . The fragmentation phenotype is similar to that observed for Atrec8 and Atmnd1 mutants, which are defective in repair of meiotic double strand breaks [34, 35]. However, since the RNAi strategy employed is not specific to meiosis, the possibility that the fragmentation phenotype may also be influenced by depletion of AtCTF7 earlier during mitosis in the progenitor cells of the meiocytes is not ruled out. Arabidopsis mutants defective in both cohesion and formation of meiotic double strand breaks do not display such fragmentation phenotypes [27, 36].
We observed severe defects in vegetative as well as reproductive growth and development, pointing to a role for AtCTF7 throughout the plant life cycle, and extending previous work showing an essential requirement for AtCTF7 in embryo development . A P AtCTF7 nlsGUS reporter was strongly expressed in root and shoot meristems, and in young buds and leaves. In young developing leaves, a polarity in expression was observed with GUS staining confined to the basal part of the leaf and absent towards the distal portion. The gradient of expression is similar to that for the cell division marker CycB1;1::GUS . Expression declined in older buds and was not observed in expanded rosette leaves. The expression of AtCTF7 is thus seen to occur in tissues that are undergoing active cell division, consistent with the known involvement of Ctf7 in promoting establishment of cohesion in conjunction with DNA replication . Within reproductive cells, expression was observed in pollen and in the female gametophyte. The sterile phenotype we observed is likely to be accounted for mainly by the defects in meiosis as well as a possible contribution from a gametophytic component.
Establishment of cohesion by Ctf7 involves acetylation of conserved lysine residues in the Smc3 subunit of cohesion which inhibits the action of the Wpl1-Pds5 complex in preventing establishment of cohesion [9, 10, 12–14]. In yeast, Drosophila, and human cells, Ctf7/Eco1 is required for the establishment of cohesion but not for association of cohesin with chromatin in interphase [5, 21, 29]. In Drosophila, a deco mutant shows reduced staining for the cohesin subunit Scc1 at prometaphase of mitosis . The strong reduction in association of AtSCC3 with chromatin in early meiotic prophase as revealed by immunostaining of meiocytes in AtCTF7-RNAi plants is similar to what has been observed for the deco mutant in prometaphase of mitosis, and suggests conservation of Ctf7/Eco1 function in plants.
In conclusion, our findings show that AtCTF7 is required for establishment of sister chromatid cohesion during meiosis in Arabidopsis, and that continued association of cohesin with chromatin in meiosis depends on AtCTF7.
Plant materials and growth conditions
The Arabidopsis thaliana strains used were of the Columbia ecotype (Col-0). Plants were grown as described in . To generate transgenic Arabidopsis, constructs were mobilized into Agrobacterium tumefaciens strain AGL-1 using triparental mating, and transformed into Arabidopsis by vacuum infiltration as described in . P AtCTF7 nlsGUS was transformed into wild-type Col, and AtCTF7-RNAi was transformed into a line carrying a P CaMV35S LhGR-N transgene . Transgenic plants were selected on MS media, containing 120 μg/ml gentamycin (Sigma-Aldrich) for AtCTF7-RNAi transformants, and 50 μg/ml kanamycin for P AtCTF7 nlsGUS transformants, and were further confirmed by PCR.
The AtCTF7 promoter, comprising 646 bp upstream of the ATG and 45 bp from AtCTF7 coding sequence, was amplified by PCR using primers Fctf7gusHindIII and Rctf7gusnlsBamH1, and cloned as a BamH1-HindIII fragment in frame with a nlsGUS tag in the pBI101.2 binary vector. For the RNAi construct, 658 bp fragments were amplified by PCR using primer pairs F1rnaiXba1 and R1rnaiBamH1, and F1rnaiXho1 and R1rnaiEcoR1, and cloned as Xba1-BamH1 and Xho1-EcoR1 fragments in opposite directions in pKANNiBAL . The RNAi cassette was excised as a XhoI-BamH1 fragment and cloned into the binary vector pZP222-6xPOP described in .
Transgenic seeds were germinated on MS plates, and grown for 7 days after which they were transferred on MS plate containing 20 μM dexamethasone (Sigma). Seedlings were analyzed for phenotypes 7 days after transfer on dexamethasone plates. Treatment of adult plants after bolting was carried out by inclusion of 20 μM dexamethasone in the watering solution which was delivered by subirrigation. Samples for meiotic analysis were collected for analysis 5 days after the start of dexamethasone treatment.
RNA isolation and quantitative RT-PCR
Total RNA was isolated using Trizol (Invitrogen) following the manufacturer’s protocol. cDNA synthesis was performed using Reverse Transcription System (Invitrogen SuperScript II) and oligo(dT) primers. Real Time PCR reactions were performed using SYBR Green PCR master mix (Applied Biosystems). GAPC was used as the internal normalization control. PCR was performed on the ABI Prism 7900 HT Sequence Detection System (Applied Biosystems) in a 384 well reaction plate according to the manufacturer’s recommendations. Primers used were Ctf7qRTF and Ctf7qRTR for AtCTF7, and GAPRTF and GAPRTR for GAPC (Additional file 1: Table S1). Cycling parameters consisted of 2 minutes incubation at 50°C, 10 minutes at 95°C, and 40 cycles of 95°C for 15 seconds, 57°C for 30 seconds and 67°C for 30 seconds. The PCR reaction was performed in triplicate for each RNA sample, and the experiment was carried out on two different biological samples representing the same RNAi line. Specificity of the amplifications was verified at the end of each PCR run using ABI prism dissociation curve analysis software. Results from the ABI Prism 7900 HT Sequence Detection System were analyzed further using Microsoft Excel. Relative amounts of mRNA were calculated from threshold points (Ct values) located in the log-linear range of real time PCR amplification plots using the 2-ΔCt method. Standard deviations in Additional file 1: Figure S1 are for variation across biological samples.
Whole mount analysis of ovules was done after fixing and clearing the inflorescence in methyl benzoate as described previously . Scanning electron microscopic (SEM) analysis of pollen was carried out using a Hitachi scanning electron microscope (model 3400 N, http://www.hitachi-hitec.com). Pollen viability was examined using Alexander staining . For DAPI analysis of pollen, anthers were squashed and stained with DAPI (1 μg/ml). Meiotic chromosome spreads were carried out as described in , with minor modifications . Observations were made on a Zeiss Axioplan 2 imaging microscope, using a Plan Apochromat 63 × oil immersion objective. Tissue from P AtCTF7 nlsGUS transgenic plants was stained for GUS activity as described in .
For FISH, chromosome spreads were carried out as described above, and FISH analysis was carried out according to the method described in , with minor modifications . The 180-bp centromeric pAL1 repeat was used to detect centromere sequences . A plasmid harboring two copies of the pAL1 repeat was subjected to PCR in the presence of Cy3-dATP (GE Healthcare), using PAL forward and reverse primers (Additional file 1: Table S1). BAC clones T19F6 and T22A6 from chromosome 4 were used as probes to monitor arm cohesion after being subjected to nick translation and labeling by Cy3-dATP (Roche). Slides were observed under a Zeiss Axioplan 2 imaging microscope equipped with a Plan Apochromat 63× oil immersion objective, using an excitation (Cy3, 550 nm) and long-pass emission (Cy3, 570 nm) filter.
For immunostaining, inflorescences were fixed as described in . Young buds were dissected out and washed with 10 mM Citrate Buffer pH4.5 (1× CB), followed by digestion with a cell wall digesting enzyme mix containing 0.3% cellulase, 0.3% pectolyase, 0.4% cytohelicase (all Sigma) in 1× CB, and incubated for 30 min at 37°C. The enzyme mix was replaced with 1× PBS, and anthers were dissected out from buds on a slide and squashed using a 22×22 mm coverslip. The slide was snap-frozen by dipping in liquid nitrogen and the coverslip was immediately removed. Slides were then dried and dipped briefly in molten 1% gelatin, 1% agarose solution to cover the cells with a thin layer of gelatin-agarose and dried. Slides were rehydrated in 1× PBS, and digested with the enzyme mix described above for 30 min at 37°C. This was followed by permeabilization of the cells in 1× PBS, 1% Triton-X100, for 30 min and washing of the slides 2–3 times in 1× PBS containing 0.1% Triton-X100. Immunostaining was performed as described in , using ASY1 antibody at a 1:1000 dilution, and AtSCC3 antibody at 1:200 dilution. All secondary antibodies were used at a dilution of 1:100. Slides were mounted in 1ug/ml DAPI in Vectashield (VectorLabs). Cells were imaged using a Zeiss Axio Imager.Z1 microscope equipped with an apotome module, using a Plan-Apochromat 63× oil-immersion objective.
While this manuscript was under review, a related study appeared online by Bolanos-Villegas et al., on the role of AtCTF7 in DNA repair, mitosis, and meiosis .
We thank Christopher Franklin and Raphael Mercier for gifts of antibodies. DS thanks S. Prabha for assistance. This work was supported by funds from the Council of Scientific and Industrial Research (CSIR), Govt. of India, and by a Centre of Excellence Grant from the Department of Biotechnology to IS. DS was supported by a fellowship from the Indian Council of Medical Research (ICMR), and SA was supported by a CSIR International Fellowship.
- Nasmyth K, Haering CH: Cohesin: Its roles and mechanisms. Annu Rev Genet. 2009, 43: 525-558. 10.1146/annurev-genet-102108-134233.PubMedView ArticleGoogle Scholar
- Rudra S, Skibbens RV: Cohesin codes - interpreting chromatin architecture and the many facets of cohesin function. J Cell Sci. 2013, 126: 31-41. 10.1242/jcs.116566.PubMedPubMed CentralView ArticleGoogle Scholar
- Nasmyth K: Cohesin: a catenase with separate entry and exit gates?. Nat Cell Biol. 2011, 13: 1170-1177. 10.1038/ncb2349.PubMedView ArticleGoogle Scholar
- Ciosk R, Shirayama M, Shevchenko A, Tanaka T, Toth A, Shevchenko A, Nasmyth K: Cohesin’s binding to chromosomes depends on a separate complex consisting of scc2 and scc4 proteins. Mol Cell. 2000, 5: 243-254. 10.1016/S1097-2765(00)80420-7.PubMedView ArticleGoogle Scholar
- Tóth A, Ciosk R, Uhlmann F, Galova M, Schleiffer A, Nasmyth K: Yeast cohesin complex requires a conserved protein, Eco1p(Ctf7), to establish cohesion between sister chromatids during DNA replication. Genes Dev. 1999, 13: 320-333. 10.1101/gad.13.3.320.PubMedPubMed CentralView ArticleGoogle Scholar
- Skibbens RV, Corson LB, Koshland D, Hieter P: Ctf7p is essential for sister chromatid cohesion and links mitotic chromosome structure to the DNA replication machinery. Genes Dev. 1999, 13: 307-319. 10.1101/gad.13.3.307.PubMedPubMed CentralView ArticleGoogle Scholar
- Moldovan G-L, Pfander B, Jentsch S: PCNA controls establishment of sister chromatid cohesion during S phase. Mol Cell. 2006, 23: 723-732. 10.1016/j.molcel.2006.07.007.PubMedView ArticleGoogle Scholar
- Sherwood R, Takahashi TS, Jallepalli PV: Sister acts: coordinating DNA replication and cohesion establishment. Genes Dev. 2010, 24: 2723-2731. 10.1101/gad.1976710.PubMedPubMed CentralView ArticleGoogle Scholar
- Sutani T, Kawaguchi T, Kanno R, Itoh T, Shirahige K: Budding yeast wpl1(rad61)-pds5 complex counteracts sister chromatid cohesion-establishing reaction. Curr Biol. 2009, 19: 492-497. 10.1016/j.cub.2009.01.062.PubMedView ArticleGoogle Scholar
- Ben-Shahar TR, Heeger S, Lehane C, East P, Flynn H, Skehel M, Uhlmann F: Eco1-dependent cohesin acetylation during establishment of sister chromatid cohesion. Science. 2008, 321: 563-566. 10.1126/science.1157774.View ArticleGoogle Scholar
- Heidinger-Pauli JM, Ünal E, Koshland D: Distinct targets of the eco1 acetyltransferase modulate cohesion in S phase and in response to DNA damage. Mol Cell. 2009, 34: 311-321. 10.1016/j.molcel.2009.04.008.PubMedPubMed CentralView ArticleGoogle Scholar
- Rowland BD, Roig MB, Nishino T, Kurze A, Uluocak P, Mishra A, Beckouët F, Underwood P, Metson J, Imre R, Mechtler K, Katis VL, Nasmyth K: Building sister chromatid cohesion: Smc3 acetylation counteracts an antiestablishment activity. Mol Cell. 2009, 33: 763-774. 10.1016/j.molcel.2009.02.028.PubMedView ArticleGoogle Scholar
- Ünal E, Heidinger-Pauli JM, Kim W, Guacci V, Onn I, Gygi SP, Koshland DE: A molecular determinant for the establishment of sister chromatid cohesion. Science. 2008, 321: 566-569. 10.1126/science.1157880.PubMedView ArticleGoogle Scholar
- Zhang J, Shi X, Li Y, Kim B-J, Jia J, Huang Z, Yang T, Fu X, Jung SY, Wang Y, Zhang P, Kim S-T, Pan X, Qin J: Acetylation of Smc3 by Eco1 is required for S phase sister chromatid cohesion in both human and yeast. Mol. Cell. 2008, 31: 143-151. 10.1016/j.molcel.2008.06.006.PubMedView ArticleGoogle Scholar
- Rudra S, Skibbens RV: Sister chromatid cohesion establishment occurs in concert with lagging strand synthesis. Cell Cycle. 2012, 11: 2114-2121. 10.4161/cc.20547.PubMedPubMed CentralView ArticleGoogle Scholar
- Song J, Lafont A, Chen J, Wu FM, Shirahige K, Rankin S: Cohesin acetylation promotes sister chromatid cohesion only in association with the replication machinery. J Biol Chem. 2012, 287: 34325-34336. 10.1074/jbc.M112.400192.PubMedPubMed CentralView ArticleGoogle Scholar
- Rivera T, Losada A: Recycling cohesin rings by deacetylation. Mol Cell. 2010, 39: 657-659. 10.1016/j.molcel.2010.08.032.PubMedView ArticleGoogle Scholar
- Beckouët F, Hu B, Roig MB, Sutani T, Komata M, Uluocak P, Katis VL, Shirahige K, Nasmyth K: An Smc3 acetylation cycle is essential for establishment of sister chromatid cohesion. Mol Cell. 2010, 39: 689-699. 10.1016/j.molcel.2010.08.008.PubMedView ArticleGoogle Scholar
- Borges V, Lehane C, Lopez-Serra L, Flynn H, Skehel M, Ben-Shahar TR, Uhlmann F: Hos1 deacetylates Smc3 to close the cohesin acetylation cycle. Mol Cell. 2010, 39: 677-688. 10.1016/j.molcel.2010.08.009.PubMedView ArticleGoogle Scholar
- Tanaka K, Yonekawa T, Kawasaki Y, Kai M, Furuya K, Iwasaki M, Murakami H, Yanagida M, Okayama H: Fission yeast eso1p is required for establishing sister chromatid cohesion during S phase. Mol Cell Biol. 2000, 20: 3459-3469. 10.1128/MCB.20.10.3459-3469.2000.PubMedPubMed CentralView ArticleGoogle Scholar
- Williams BC, Garrett-Engele CM, Li Z, Williams EV, Rosenman ED, Goldberg ML: Two putative acetyltransferases, San and Deco, are required for establishing sister chromatid cohesion in Drosophila. Curr Biol. 2003, 13: 2025-2036. 10.1016/j.cub.2003.11.018.PubMedView ArticleGoogle Scholar
- Vega H, Waisfisz Q, Gordillo M, Sakai N, Yanagihara I, Yamada M, van Gosliga D, Kayserili H, Xu C, Ozono K, Jabs EW, Inui K, Joenje H: Roberts syndrome is caused by mutations in Esco2, a human homolog of yeast Eco1 that is essential for the establishment of sister chromatid cohesion. Nat Genet. 2005, 37: 468-470. 10.1038/ng1548.PubMedView ArticleGoogle Scholar
- Jiang L, Yuan L, Xia M, Makaroff CA: Proper levels of the Arabidopsis cohesion establishment factor Ctf7 are essential for embryo and megagametophyte, but not endosperm, Development. Plant Physiol. 2010, 154: 820-832. 10.1104/pp.110.157560.PubMedPubMed CentralView ArticleGoogle Scholar
- Sebastian J, Ravi M, Andreuzza S, Panoli AP, Marimuthu MPA, Siddiqi I: The plant adherin AtSCC2 is required for embryogenesis and sister-chromatid cohesion during meiosis in Arabidopsis. Plant J. 2009, 59: 1-13. 10.1111/j.1365-313X.2009.03845.x.PubMedView ArticleGoogle Scholar
- Craft J, Samalova M, Baroux C, Townley H, Martinez A, Jepson I, Tsiantis M, Moore I: New pOp/LhG4 vectors for stringent glucocorticoid-dependent transgene expression in Arabidopsis. Plant J. 2005, 41: 899-918. 10.1111/j.1365-313X.2005.02342.x.PubMedView ArticleGoogle Scholar
- Schneitz K, Hülskamp M, Pruitt RE: Wild-type ovule development in Arabidopsis thaliana: a light microscope study of cleared whole-mount tissue. Plant J. 1995, 7: 731-749. 10.1046/j.1365-313X.1995.07050731.x.View ArticleGoogle Scholar
- Chelysheva L, Diallo S, Vezon D, Gendrot G, Vrielynck N, Belcram K, Rocques N, Márquez-Lema A, Bhatt AM, Horlow C, Mercier R, Mézard C, Grelon M: AtREC8 and AtSCC3 are essential to the monopolar orientation of the kinetochores during meiosis. J Cell Sci. 2005, 118: 4621-4632. 10.1242/jcs.02583.PubMedView ArticleGoogle Scholar
- Armstrong SJ, Caryl AP, Jones GH, Franklin FCH: Asy1, a protein required for meiotic chromosome synapsis, localizes to axis-associated chromatin in Arabidopsis and Brassica. J Cell Sci. 2002, 115: 3645-3655. 10.1242/jcs.00048.PubMedView ArticleGoogle Scholar
- Hou F, Zou H: Two human orthologues of Eco1/Ctf7 acetyltransferases are both required for proper sister-chromatid cohesion. Mol Biol Cell. 2005, 16: 3908-3918. 10.1091/mbc.E04-12-1063.PubMedPubMed CentralView ArticleGoogle Scholar
- Guacci V, Koshland D: Cohesin-independent segregation of sister chromatids in budding yeast. Mol Biol Cell. 2012, 23: 729-739. 10.1091/mbc.E11-08-0696.PubMedPubMed CentralView ArticleGoogle Scholar
- Ghosh S, Gardner JM, Smoyer CJ, Friederichs JM, Unruh JR, Slaughter BD, Alexander R, Chisholm RD, Lee KK, Workman JL, Jaspersen SL: Acetylation of the SUN protein Mps3 by Eco1 regulates its function in nuclear organization. Mol Biol Cell. 2012, 23: 2546-2559. 10.1091/mbc.E11-07-0600.PubMedPubMed CentralView ArticleGoogle Scholar
- Evans EB, Hogarth C, Evanoff RM, Mitchell D, Small C, Griswold MD: Localization and regulation of murine Esco2 during male and female meiosis. Biol Reprod. 2012, 87: 61-10.1095/biolreprod.112.099978.PubMedPubMed CentralView ArticleGoogle Scholar
- Lu S, Goering M, Gard S, Xiong B, McNairn AJ, Jaspersen SL, Gerton JL: Eco1 is important for DNA damage repair in S cerevisiae. Cell Cycle. 2010, 9: 3315-3327. 10.4161/cc.9.16.12673.PubMedPubMed CentralGoogle Scholar
- Bai X, Peirson BN, Dong F, Xue C, Makaroff CA: Isolation and characterization of SYN1, a RAD21-like gene essential for meiosis in Arabidopsis. Plant Cell. 1999, 11: 417-430.PubMedPubMed CentralView ArticleGoogle Scholar
- Panoli AP, Ravi M, Sebastian J, Nishal B, Reddy TV, Marimuthu MP, Subbiah V, Vijaybhaskar V, Siddiqi I: AtMND1 is required for homologous pairing during meiosis in Arabidopsis. BMC Mol Biol. 2006, 7: 24-10.1186/1471-2199-7-24.PubMedPubMed CentralView ArticleGoogle Scholar
- Mercier R, Armstrong SJ, Horlow C, Jackson NP, Makaroff CA, Vezon D, Pelletier G, Jones GH, Franklin FCH: The meiotic protein SWI1 is required for axial element formation and recombination initiation in Arabidopsis. Development. 2003, 130: 3309-3318. 10.1242/dev.00550.PubMedView ArticleGoogle Scholar
- Donnelly PM, Bonetta D, Tsukaya H, Dengler RE, Dengler NG: Cell cycling and cell enlargement in developing leaves of Arabidopsis. Dev Biol. 1999, 215: 407-419. 10.1006/dbio.1999.9443.PubMedView ArticleGoogle Scholar
- Siddiqi I, Ganesh G, Grossniklaus U, Subbiah V: The dyad gene is required for progression through female meiosis in Arabidopsis. Development. 2000, 127: 197-207.PubMedGoogle Scholar
- Bechtold N, Pelletier G: In planta Agrobacterium-mediated transformation of adult Arabidopsis thaliana plants by vacuum infiltration. Methods Mol Biol. 1998, 82: 259-266.PubMedGoogle Scholar
- Wesley SV, Helliwell CA, Smith NA, Wang M, Rouse DT, Liu Q, Gooding PS, Singh SP, Abbott D, Stoutjesdijk PA, Robinson SP, Gleave AP, Green AG, Waterhouse PM: Construct design for efficient, effective and high-throughput gene silencing in plants. Plant J. 2001, 27: 581-590. 10.1046/j.1365-313X.2001.01105.x.PubMedView ArticleGoogle Scholar
- Alexander MP: Differential staining of aborted and nonaborted pollen. Stain Technol. 1969, 44: 117-122.PubMedGoogle Scholar
- Ross KJ, Fransz P, Jones GH: A light microscopic atlas of meiosis in Arabidopsis thaliana. Chromosome Res. 1996, 4: 507-516. 10.1007/BF02261778.PubMedView ArticleGoogle Scholar
- Agashe B, Prasad CK, Siddiqi I: Identification and analysis of DYAD: a gene required for meiotic chromosome organisation and female meiotic progression in Arabidopsis. Development. 2002, 129: 3935-3943.PubMedGoogle Scholar
- Fransz P, Armstrong S, Alonso-Blanco C, Fischer TC, Torres-Ruiz RA, Jones G: Cytogenetics for the model system Arabidopsis thaliana. Plant J. 1998, 13: 867-876. 10.1046/j.1365-313X.1998.00086.x.PubMedView ArticleGoogle Scholar
- Martinez-Zapater JM, Estelle MA, Somerville CR: A highly repeated DNA sequence in Arabidopsis thaliana. Mol Gen Genet. 1986, 204: 417-423. 10.1007/BF00331018.View ArticleGoogle Scholar
- Cai X, Dong F, Edelmann RE, Makaroff CA: The Arabidopsis SYN1 cohesin protein is required for sister chromatid arm cohesion and homologous chromosome pairing. J Cell Sci. 2003, 116: 2999-3007. 10.1242/jcs.00601.PubMedView ArticleGoogle Scholar
- Bolaños-Villegas P, Yang X, Wang HJ, Juan CT, Chuang MH, Makaroff CA, Jauh GY: Arabidopsis Chromosome Transmission Fidelity 7 (AtCTF7/ECO1) is required for DNA repair, mitosis and meiosis. Plant J. 2013, 10.1111/tpj.12261.Google Scholar