The ARABIDOPSIS SKP1-LIKE1 (ASK1) protein acts predominately from leptotene to pachytene and represses homologous recombination in male meiosis

Abstract

Normal progression of genetic recombination requires timely degradation of many proteins, but little is known about the proteolytic mechanism. The ARABIDOPSIS SKP1-LIKE1 (ASK1) protein is a component of the Skp1-Cullin-F-box-protein (SCF) ubiquitin ligases that target a variety of proteins for degradation via the 26S proteasome pathway. Previous studies indicate that the early defects of the mutant ask1-1 occur in a prophase-I period overlapping with the period of homologous recombination. We provide evidence in this report that ASK1 is predominately expressed from leptotene to pachytene, and negatively regulates recombination. First, the ASK1 transcript was found not to co-exist with that of its closest homolog ASK2 only during prophase I of male meiosis, suggesting that ASK1 is functionally non-redundant only in prophase I. Second, the peak level of an ASK1-green fluorescence protein (GFP) fusion protein expressed by an ASK1 promoter region occurred only from leptotene to pachytene. The ASK1-GFP in a dominant negative fashion resulted in abnormal tetrads resembling those of the ask1-1 mutant, supporting that the expression timing of the ASK1-GFP in male meiocytes reflects the expression timing of the endogenous ASK1. Lastly, using a marker for recombination events, a significant increase in recombination frequency was detected in plants heterozygous for ask1-1. These results indicate that ASK1 normally plays a repressive role in male recombination in Arabidopsis.