BBX21 (also known as SALT TOLERANCE HOMOLOG 2), a B-box (BBX)-containing protein, has been previously identified as a positive regulator of light signaling; however, the precise role of BBX21 in regulating seedling photomorphogenesis remains largely unclear. In this study, we report that CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1) interacts with BBX21 in vivo and is able to ubiquitinate BBX21 in vitro. Thus, BBX21 is targeted for 26S proteasome-mediated degradation in dark-grown Arabidopsis seedlings in a COP1-dependent manner. Moreover, we show that BBX21 binds to the T/G-box in the ELONGATED HYPOCOTYL 5 (HY5) promoter and directly activates HY5 expression in the light. Transgenic seedlings overexpressing BBX21 exhibit dramatically shortened hypocotyls in the light, and this phenotype is dependent on a functional HY5. Taken together, our data suggest a molecular base underlying BBX21-mediated seedling photomorphogenesis, indicating that BBX21 is a pivotal component involved in the COP1-HY5 regulatory hub.P lant seedlings undergo two distinct developmental processes dependent on the presence and absence of light, termed photomorphogenesis and skotomorphogenesis, respectively (1). CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1) is a central repressor of seedling photomorphogenesis, and mutants impaired in COP1 display constitutively photomorphogenic phenotypes in darkness (2, 3). COP1 acts as an E3 ubiquitin ligase targeting a subset of substrates for degradation in darkness, including ELONGATED HYPOCOTYL 5 (HY5), HY5 HOMOLOG (HYH), LONG AFTER FAR-RED LIGHT 1 (LAF1), LONG HYPOCOTYL IN FAR RED (HFR1), B-BOX PROTEIN 22/ SALT TOLERANCE HOMOLOG 3 (BBX22/STH3), and PHYTOCHROME INTERACTING FACTOR 3-LIKE1 (PIL1) (4-11). Of these, HY5 is considered a key signal integration point from dark to light transition (10, 12, 13). Its abundance is directly correlated with the extent of seedling photomorphogenic growth, but inversely correlated with the nuclear abundance of COP1 (4, 10, 14). As a b-ZIP type transcription factor, HY5 specifically interacts with the ACGT-containing elements (ACEs) and might directly bind to approximately one-third of the Arabidopsis genes (12, 13). Thus, HY5 ensures proper expression of a large number of downstream regulatory genes, which in turn eventually promotes photomorphogenesis in the light. Recent studies have shown that HY5 expression is regulated by HY5 itself, as well as by HYH and CALMODULIN7 (CAM7) (15,16).A total of 32 B-box (BBX)-containing proteins have been identified in the Arabidopsis genome (17), among which a number of BBX proteins have been shown to be involved in COP1-and HY5-mediated seedling photomorphogenesis (18). BBX4, BBX20, BBX22, BBX24, and BBX25 genetically and physically interact with COP1 and undergo COP1-mediated degradation in the dark (6,(19)(20)(21)(22)(23)(24). In addition, BBX22, BBX24, and BBX25 physically interact with HY5, and, interestingly, BBX22 acts as a coactivator of HY5 action (6), whereas BBX24 and BBX25 repress the transcriptional activating activity of HY5 ...

Light is one of the most important environmental cues regulating multiple aspects of plant growth and development, and abscisic acid (ABA) is a plant hormone that plays important roles during many phases of the plant life cycle and in plants' responses to various environmental stresses. How plants integrate the external light signal with endogenous ABA pathway for better adaptation and survival remains poorly understood. Here, we show that BBX21 (also known as SALT TOLERANCE HOMOLOG 2), a B-box (BBX) protein previously shown to positively regulate seedling photomorphogenesis, is also involved in ABA signaling. Our genetic data show that BBX21 may act upstream of several ABA INSENSITIVE (ABI) genes and ELONGATED HYPOCOTYL 5 (HY5) in ABA control of seed germination. Previous studies showed that HY5 acts as a direct activator of ABI5 expression, and that BBX21 interacts with HY5. We further demonstrate that BBX21 negatively regulates ABI5 expression by interfering with HY5 binding to the ABI5 promoter. In addition, ABI5 was shown to directly activate its own expression, whereas BBX21 negatively regulates this activity by directly interacting with ABI5. Together, our study indicates that BBX21 coordinates with HY5 and ABI5 on the ABI5 promoter and that these transcriptional regulators work in concert to integrate light and ABA signaling in Arabidopsis thaliana.

Seedling photomorphogenesis is a sophisticated developmental process that is controlled by both the transcriptional and posttranscriptional regulation of gene expression. Here, we identify an Arabidopsis noncoding RNA, designated HIDDEN TREASURE 1 (HID1), as a factor promoting photomorphogenesis in continuous red light (cR). We show that HID1 acts through PHYTOCHROME-INTERACTING FACTOR 3 (PIF3), which encodes a basic helix-loophelix transcription factor known to be a key repressor of photomorphogenesis. Knockdown of HID1 in hid1 mutants leads to a significant increase in the expression of PIF3, which in turn drives the development of elongated hypocotyls in cR. We identified two major stem-loops in HID1 that are essential for its modulation of hypocotyl growth in cR-grown seedlings. Furthermore, our data reveal that HID1 is assembled into large nuclear protein-RNA complex(es) and that it associates with the chromatin of the first intron of PIF3 to repress its transcription. Strikingly, phylogenetic analysis reveals that many land plants have conserved homologs of HID1 and that its rice homolog can rescue the mutant phenotype when expressed in Arabidopsis hid1 mutants. We thus concluded that HID1 is a previously uncharacterized noncoding RNA whose function represents another layer of regulation in the precise control of seedling photomorphogenesis.light signaling | transcriptional regulation