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The rice U-box/ARM E3 ubiquitin ligase SPL11 negatively regulates programmed cell death (PCD) and disease resistance, and controls flowering time through interacting with the novel RNA/DNA binding KH domain protein SPIN1. Overexpression of  Spin1  causes late flowering in transgenic rice under short-day (SD) and long-day (LD) conditions. In this study, we characterized the function of the  R NA- b inding and  S PIN1-interacting  1  (RBS1) protein in flowering time regulation.  Rbs1 was identified in a yeast-two-hybrid screen using the full-length  Spin1  cDNA as a bait and encodes an RNA binding protein with three RNA recognition motifs. The protein binds RNA  in vitro  and interacts with SPIN1 in the nucleus.  Rbs1  overexpression causes delayed flowering under SD and LD conditions in rice. Expression analyses of flowering marker genes show that  Rbs1  overexpression represses the expression of  Hd3a  under SD and LD conditions.  Rbs1  is upregulated in both  Spin1  overexpression plants and in the  spl11  mutant. Interestingly,  Spin1  expression is increased but  Spl11  expression is repressed in the  Rbs1  overexpression plants. Western blot analysis revealed that the SPIN1 protein level is increased in the  Rbs1  overexpression plants and that the RBS1 protein level is also up-regulated in the  Spin1  overexpression plants. These results suggest that RBS1 is a new negative regulator of flowering time that itself is positively regulated by SPIN1 but negatively regulated by SPL11 in rice.

RBS1, an RNA Binding Protein, Interacts with SPIN1 and Is Involved in Flowering Time Control in Rice