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. 2018 Jan 30;176(4):3136–3145. doi: 10.1104/pp.17.01743

Figure 4.

Figure 4.

Inhibition of CRY1 by the synthetic microProtein miP-CRY1. A, Domain structures of CRY1 and miP-CRY1. B, Model depicting CRY1 function and miP-CRY1 inhibition. The CRY1 homodimer is activated by a blue light signal. Activated CRY1 regulates de-etiolation of the plant. MiP-CRY1 forms a nonfunctional heterodimer with CRY1 that leads to hypocotyl elongation under blue light. C, Image of representative Col-0, cry1cry2, and two independent 35S::miP-CRY1 lines grown under white light, dark, or blue light conditions. D, RT-qPCR showing expression of endogenous CRY1 and transgene miP-CRY1 in 35S::miP-CRY1 lines compared with Col-0. Experimental replicates were statistically tested using a Student’s t test (**P < 0.01). E, Boxplot of hypocotyl length under white light, dark, and blue light (n = 10; *P < 0.05, **P < 0.01, t test). F, Interaction of miP-CRY1 with CRY1 was tested using a yeast two-hybrid assay. Normal yeast growth was observed for the serial dilutions on nonselective SD medium lacking Leu and Trp (SD-L-W). Only positive interactors grew on selective SD medium lacking Leu, Trp, and His (SD-L-W-H) supplemented with 10 mm 3-aminotriazole (3-AT).