Ahp2 (Hop2) function in Arabidopsis thaliana (Ler) is required for stabilization of close alignment and synaptonemal complex formation except for the two short arms that contain nucleolus organizer regions.
A cytological comparative analysis of male meiocytes was performed for Arabidopsis wild type and the ahp2 (hop2) mutant with emphasis on ahp2's largely uncharacterized prophase I. Leptotene progression appeared normal in ahp2 meiocytes; chromosomes exhibited regular axis formation and assumed a typical polarized nuclear organization. In contrast, 4',6'-diamidino-2-phenylindole-stained ahp2 pachytene chromosome spreads demonstrated a severe reduction in stabilized pairing. However, transmission electron microscopy (TEM) analysis of sections from meiocytes revealed that ahp2 chromosome axes underwent significant amounts of close alignment (44% of total axis). This apparent paradox strongly suggests that the Ahp2 protein is involved in the stabilization of homologous chromosome close alignment. Fluorescent in situ hybridization in combination with Zyp1 immunostaining revealed that ahp2 mutants undergo homologous synapsis of the nucleolus-organizer-region-bearing short arms of chromosomes 2 and 4, despite the otherwise "nucleus-wide" lack of stabilized pairing. The duration of ahp2 zygotene was significantly prolonged and is most likely due to difficulties in chromosome alignment stabilization and subsequent synaptonemal complex formation. Ahp2 and Mnd1 proteins have previously been shown, "in vitro," to form a heterodimer. Here we show, "in situ," that the Ahp2 and Mnd1 proteins are synchronous in their appearance and disappearance from meiotic chromosomes. Both the Ahp2 and Mnd1 proteins localize along the chromosomal axis. However, localization of the Ahp2 protein was entirely foci-based whereas Mnd1 protein exhibited an immunostaining pattern with some foci along the axis and a diffuse staining for the rest of the chromosome.
Stronghill
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Department of Cell and Systems Biology, University of Toronto, Scarborough, Ontario, Canada. stronghill@utsc.utoronto.ca
Pathan
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Supijono
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Hasenkampf
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D000705:Anaphase; D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D023902:Chromosome Pairing; D032461:Chromosomes, Plant / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D017404:In Situ Hybridization, Fluorescence; D008540:Meiosis; D049469:Meiotic Prophase I; D046529:Microscopy, Electron, Transmission; D009697:Nucleolus Organizer Region / Q000502:physiology*; D010770:Phosphotransferases / Q000235:genetics / Q000378:metabolism*; D013573:Synaptonemal Complex / Q000235:genetics / Q000378:metabolism*; D000097722:Cohesins
D029681:Arabidopsis Proteins; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D010770:Phosphotransferases; D000097722:Cohesins; C117967:AHP2 protein, Arabidopsis; C514180:MND1 protein, Arabidopsis