Activation of the Arabidopsis membrane-bound transcription factor bZIP28 is mediated by site-2 protease, but not site-1 protease.

The unfolded protein response (UPR) is a homeostatic cellular response conserved in eukaryotic cells to alleviate the accumulation of unfolded proteins in the endoplasmic reticulum (ER). Arabidopsis bZIP28 is a membrane-bound transcription factor activated by proteolytic cleavage in response to ER stress, thereby releasing its cytosolic portion containing the bZIP domain from the membrane to translocate into the nucleus where it induces the transcription of genes encoding ER-resident molecular chaperones and folding enzymes. It has been widely recognized that the proteolytic activation of bZIP28 is mediated by the sequential cleavage of site-1 protease (S1P) and site-2 protease (S2P). In the present study we provide evidence that bZIP28 protein is cleaved by S2P, but not by S1P. We demonstrated that wild-type and s1p mutant plants produce the active, nuclear form of bZIP28 in response to the ER stress inducer tunicamycin. In contrast, tunicamycin-treated s2p mutants do not accumulate the active, nuclear form of bZIP28. Consistent with these observations, s2p mutants, but not s1p mutants, exhibited a defective transcriptional response of ER stress-responsive genes and significantly higher sensitivity to tunicamycin. Interestingly, s2p mutants accumulate two membrane-bound bZIP28 fragments with a shorter ER lumen-facing C-terminal domain. Importantly, the predicted cleavage sites are located far from the canonical S1P recognition motif previously described. We propose that ER stress-induced proteolytic activation of bZIP28 is mediated by the sequential actions of as-yet-unidentified protease(s) and S2P, and does not require S1P.

Exogenous salicylic acid activates two signaling arms of the unfolded protein response in Arabidopsis.

The unfolded protein response (UPR) is a highly conserved cellular response that prevents abnormal maturation of proteins in the endoplasmic reticulum (ER). The expression of genes encoding ER chaperones is induced during the UPR. In the Arabidopsis UPR, two membrane-bound transcription factors, bZIP60 and bZIP28, activate the expression of those genes. bZIP60 is regulated by unconventional cytoplasmic splicing catalyzed by inositol requiring enzyme 1 (IRE1), which is located in the ER membrane. bZIP28 is regulated by intramembrane proteolysis. Pathogen infection and salicylic acid (SA) have been reported to induce the expression of some UPR genes. Here, we show that UPR genes including BiP3, a marker gene of the Arabidopsis UPR, are induced by exogenous SA treatment and activation of bZIP60 in an IRE1-dependent manner. The induction of gene expression and activation of bZIP60 were independent of NPR1 and HsfB1 under these experimental conditions. We generated antibodies to detect the proteolytic products of bZIP28 after SA treatment. An assay using these antibodies showed that SA activated bZIP28, as well as activating bZIP60 through IRE1. Together, these results show that exogenous SA treatment activates two signaling arms of the Arabidopsis UPR. We propose a possible mechanism of activation of the UPR machinery by SA.

[A clinical analysis of patients with early-stage prostate cancer managed without initial treatment].

To search for a more suitable qualification indicating watchful waiting, we performed a retrospective study against early-stage prostate cancer patients managed without initial treatment. Thirty-three patients who had not been treated for more than 6 months after diagnosed as T1c or T2 prostate cancer were studied. The median values of total observation period, age at diagnosis, and initial PSA were 27.0 months, 69.0 years old, and 7.0 ng/ml, respectively. Among 28 patients who had had measurement of serum PSA at least three times, seven patients showed a significant PSA elevation when transition of PSA level was analyzed using linear regression analysis. The other patients had been stable or PSA level declined. Between these two groups, there was no significant difference regarding age, initial PSA, PSA density, Gleason score, number of cancer-positive core, and cancer-occupying rate in biopsy specimen. The median PSA doubling time in patients showing PSA elevation was 36.3 months. There were no patients showing PSA elevation among those with a cancer-occupying rate of less than 5%. Clinical disease progression was obviously observed in two cases although one did not show PSA elevation. During the observation period, treatment was eventually started in seven patients. The 5-year rate of no treatment was 53.8%. Although a significant independent factor predicting the future treatment was not identified, univariate analysis revealed that the initial PSA value in patients undergoing treatment was significantly higher than that of those without treatment (p = 0.032). We concluded that early-stage prostate cancer has clinical variability, and regular clinical evaluations as digital rectal examination should be performed when the patient was managed with watchful waiting.