The rice E3 ubiquitin ligase-transcription factor module targets two trypsin inhibitors to enhance broad-spectrum disease resistance.

Broad-spectrum disease resistance (BSR) is crucial for controlling plant diseases and relies on immune signals that are subject to transcriptional and post-translational regulation. How plants integrate and coordinate these signals remains unclear. We show here that the rice really interesting new gene (RING)-type E3 ubiquitin ligase OsRING113 targets APIP5, a negative regulator of plant immunity and programmed cell death (PCD), for 26S proteasomal degradation. The osring113 mutants in Nipponbare exhibited decreased BSR, while the overexpressing OsRING113 plants showed enhanced BSR against Magnaporthe oryzae (M. oryzae) and Xanthomonas oryzae pv. oryzae (Xoo). Furthermore, APIP5 directly suppressed the transcription of the Bowman-Birk trypsin inhibitor genes OsBBTI5 and AvrPiz-t-interacting protein 4 (APIP4). Overexpression of these two genes, which are partially required for APIP5-mediated PCD and disease resistance, conferred BSR. OsBBTI5 and APIP4 associated with and stabilized the pathogenesis-related protein OsPR1aL, which promotes M. oryzae resistance. Our results identify an immune module with integrated and coordinated hierarchical regulations that confer BSR in plants.

An ORFeome of rice E3 ubiquitin ligases for global analysis of the ubiquitination interactome.

BACKGROUND: Ubiquitination is essential for many cellular processes in eukaryotes, including 26S proteasome-dependent protein degradation, cell cycle progression, transcriptional regulation, and signal transduction. Although numerous ubiquitinated proteins have been empirically identified, their cognate ubiquitin E3 ligases remain largely unknown. RESULTS: Here, we generate a complete ubiquitin E3 ligase-encoding open reading frames (UbE3-ORFeome) library containing 98.94% of the 1515 E3 ligase genes in the rice (Oryza sativa L.) genome. In the test screens with four known ubiquitinated proteins, we identify both known and new E3s. The interaction and degradation between several E3s and their substrates are confirmed in vitro and in vivo. In addition, we identify the F-box E3 ligase OsFBK16 as a hub-interacting protein of the phenylalanine ammonia lyase family OsPAL1-OsPAL7. We demonstrate that OsFBK16 promotes the degradation of OsPAL1, OsPAL5, and OsPAL6. Remarkably, we find that overexpression of OsPAL1 or OsPAL6 as well as loss-of-function of OsFBK16 in rice displayed enhanced blast resistance, indicating that OsFBK16 degrades OsPALs to negatively regulate rice immunity. CONCLUSIONS: The rice UbE3-ORFeome is the first complete E3 ligase library in plants and represents a powerful proteomic resource for rapid identification of the cognate E3 ligases of ubiquitinated proteins and establishment of functional E3-substrate interactome in plants.

[Influence of Combination with 1q21 Amplification or-No in Patients with Newly Diagnosed MM on the Clinical Effecacy of Bortezomib-based induction chemotherapy and Long-Term Prognosis of Patients].

OBJECTIVE: To investigate the influence of influence of combination with 1q21 amplification or-no in patients with newly diagnosed MM on the clinical effecacy of bortezomib-based induction chemotherapy and long-term prognosis of patients. METHODS: 148 patients with newly diagnosed MM treated from January 2010 to May 2018 were selected and divided into 2 groups: group A (70 patients) without 1q21 amplification and group B (78 patients) with 1q21 amplification; and the survival benefit and influence on clinical efficacy of bortezomib were compared between 2 groups, and the factors influencing clinical prognosis in the patients with newly diagnosed MM were analyzed. RESULTS: The median PFS and OS of patients in B group were significantly shorter than those in group A (P＜0.05). There was no significant difference in the median OS and PFS between patients with 1q21 amplification copies number =3 and ＞3 (P＞0.05). Multivariate Cox model analysis indicated that the adverse factors for OS were ISS staging, Hb levels, ß2 microglobulin levels and 1q21 amplification respectively, and the adverse factors for PFS were Hb levels and 1q21 amplification respectively in patients with newly diagnosed MM (P＜0.05). The very good partial remission rate of newly diagnosed MM patients with 1q21 amplification and bortezomib-based induction chemotherapy were significantly higher than that in the patients without bortezomib-based induction chemotherapy (P＜0.05). The median PFS time of newly diagnosed MM patients with 1q21 amplification and auto-HSCT after bortezomib-based induction chemotherapy was significantly longer than that of patients without auto-HSCT (P＜0.05). CONCLUSION: 1q21 amplification should be the independent risk factor for poor prognosis of patients with newly treated MM. The application of bortezomib-containing induction chemotherapy in patients with 1q21 amplification can efficiently improve the remission rate, while auto-HSCT consolidation therapy may prolong patients' PFS.

[Biomechanical characteristics analysis on discs with coflex fixation on the different segments of lower lumbar spine].

OBJECTIVE: To investigate the biomechanical differences between the surgery and adjacent segments of intervertebral discs in the lower lumbar spine, which were implanted with Coflex into the segments of L4, and L5S1, respectively. METHODS: Three finite-element models (the model of the intact lower lumbar sacrum,the L4.5 and L5S1 segments implanted by Coflex) were developed, respectively. According to the spinal three-column loading theory, three models were forced by the physiological loads of upright standing, flexion and extension. The stress of the different areas of the disc annulus, the changes of intervertebral dorsal height and the degree of nucleus pulposus pressure were compared and analyzed. RESULTS: Coflex implanted into the L4.5 and L5S1 segments in compression and extension could both decrease the stress of the posterior area of intervertebral disc in the surgery segment, resist the changes of the intervertebral disc dorsal height and reduce the perssure of nucleus pulposus. Furthermore, the stress of the L5S1 segment decreased when Coflex fixed the L4.5 segment in extension. However, when Coflex fixed the L5S1 segment, the stress of L4.5 segment had no significant changes. CONCLUSION: Coflex fixing the L4,5 and L5S1 segments can effectively decrease the stress of the surgery segmental discs, respectively. Furthermore, Coflex fixing L4,5 segment may play a biomechanical role in reducing the stress of L5S1 segment.