Mutations in Arabidopsis condensin genes disrupt embryogenesis, meristem organization and segregation of homologous chromosomes during meiosis.
Proper chromatin condensation and sister chromatid resolution are essential for the maintenance of chromosomal integrity during cell division, and is in part mediated by a conserved multisubunit apparatus termed the condensin complex. The core subunits of the complex are members of the SMC2 (Structural Maintenance of Chromosomes) and SMC4 gene families. We have cloned an Arabidopsis gene, AtCAP-E1, which is a functional ortholog of the yeast SMC2 gene. A second, highly homologous SMC2 gene, AtCAPE-2, was identified by the Arabidopsis genome project. SMC2 gene expression in Arabidopsis was correlated with the mitotic activity of tissues, with high level expression observed in meristematic cells. The two genes are differentially expressed with AtCAP-E1 accounting for more than 85% of the total SMC2 transcript pool. The titan3 mutant is the result of a T-DNA insertion into AtCAP-E1, but other than subtle endosperm defects, titan3 is viable and fecund. We identified a T-DNA insertion mutant of AtCAP-E2, which showed no obvious mutant phenotype, indicating that the two genes are functionally redundant. Genetic crosses were employed to examine the consequences of reduced SMC2 levels. Both male and female gametogenesis were compromised in double mutant spores. Embryo lethality was observed for both double homozygous and AtCAP-E1(-/-), AtCAP-E2(+/-) plants; arrest occurred at or before the globular stage and was associated with altered planes of cell division in both the suspensor and the embryo. Down regulation of both genes by antisense technology, as well as in AtCAP-E1(+/-), AtCAP-E2(-/-) plants results in meristem disorganization and fasciation. Our data are consistent with the interpretation that threshold levels of SMC2 proteins are required for normal development and that AtCAP-E2 may have a higher affinity for its target than AtCAP-E1.
Siddiqui
Najeeb U
NU
Department of Botany, Division of Life Sciences, University of Toronto, 1265 Military Trail, West Hill, Ontario M1C 1A4, Canada.
Stronghill
Patricia E
PE
Dengler
Ronald E
RE
Hasenkampf
Clare A
CA
Riggs
C Daniel
CD
D000251:Adenosine Triphosphatases / Q000235:genetics* / Q000502:physiology*; D000483:Alleles; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000502:physiology*; D002843:Chromatin / Q000378:metabolism; D003001:Cloning, Molecular; D003433:Crosses, Genetic; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000502:physiology*; D017343:Genes, Plant; D005816:Genetic Complementation Test; D018745:Genome, Plant; D006720:Homozygote; D015151:Immunoblotting; D017403:In Situ Hybridization; D008540:Meiosis; D018519:Meristem / Q000502:physiology*; D008856:Microscopy, Fluorescence; D008938:Mitosis; D008957:Models, Genetic; D046912:Multiprotein Complexes; D009154:Mutation; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010641:Phenotype; D018521:Plant Physiological Phenomena; D030821:Plants, Genetically Modified; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors
D029681:Arabidopsis Proteins; D002843:Chromatin; D004268:DNA-Binding Proteins; D046912:Multiprotein Complexes; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; C108312:condensin complexes; D000251:Adenosine Triphosphatases