Substitution of a Conserved Glycine in the PHR Domain of Arabidopsis CRYPTOCHROME 1 Confers a Constitutive Light Response

Abstract

CRYPTOCHROMES (CRYs) are photolyase-like ultraviolet-A/blue light photoreceptors that mediate various light responses in plants. The signaling mechanism of Arabidopsis CRYs (CRY1 and CRY2) involves direct CRY-COP1 interaction. Here, we report that CRY1(G380R), which carries a Gly-to-Arg substitution of the highly conserved G380 in the photolyase-related (PHR) domain of Arabidopsis CRY1, shows constitutive CRY1 photoreceptor activity in Arabidopsis. Transgenic plants overexpressing CRY1(G380R) display a constitutively photomorphogenic (COP) phenotype in darkness, as well as a dramatic early flowering phenotype under short-day light conditions (SD). We further demonstrate that CRY1(G380R) expression driven by the native CRY1 promoter also results in a COP phenotype in darkness. Moreover, overexpression of either the Arabidopsis homolog CRY2(G377R) or the rice ortholog OsCRY1b(G388R) of CRY1(G380R) in Arabidopsis results in a COP phenotype in darkness. Cellular localization studies indicate that CRY1(G380R) co-localizes with COP1 in the same nuclear bodies (NBs) in vivo and inhibits the nuclear accumulation of COP1 in darkness. These results suggest that the conserved G380 may play a critical role in regulating the photoreceptor activity of plant CRYs and that CRY1(G380R) might constitutively phenocopy the photo-activated CRY1 in darkness and thus constitutively mediate CRY1 signaling.