Coordinated Expression of Cytosolic and Chloroplastic Glutamine Synthetase During Reproductive Stage and Its Impact in GS1 RNAi Transgenic Rice

To understand the reallocation of organic nitrogen from leaf to the flower head of rice, the role of glutamine synthetase （GS） was investigated by characterizing GS1 RNAi transgenic rice, which revealed a significant reduction in panicle number and number of seeds per panicle. We observed the expression of GS isotypes at transcriptional and protein levels in flag leaves, leaf sheaths and panicles at three different flower development stages. The mRNA expression of GS1;1 was clearly suppressed in flag leaves, especially at the flowering stage. GS1 protein was barely detectable in flag leaves until the flowering stage, while GS1 protein was compromised in the leaf sheath and panicle, with transient expression of GS2 protein at the flowering stage. The glutamine level in transgenic plants was significantly reduced in both flag leaves and panicles, but ammonium was highly accumulated. The level of other amino acids, including aspartate and asparagine, tended to be higher in RNAi transgenic plants than the wild type plants during the reproductive stage. In addition, accumulation of toxic ammonium in panicles with low glutamine level might have caused low seed-setting in the transgenic rice. These results indicated that nitrogen reallocation was critical for panicle development, and that multiple GS isotypes functioned cooperatively to complete the rice life cycle when leaf nitrogen was remobilized to the developing reproductive organs.

pigment epithelium-derived factor protects the morphological structure of retinal Müller cells in diabetic rats

AIM: To investigate if pigment epithelium-derived factor(PEDF) has any protective effect on the retinal Müller cells of Sprague-Dawley rats suffering from diabetes mellitus.METHODS: Sixty Sprague-Dawley rats were randomly divided into a negative control group, a group receiving0.1 μg/μL PEDF, another group receiving 0.2 μg/μL PEDF,and a group receiving balanced salt solution(BSS). Rats in both the PEDF and BSS groups were treated intravitreally based on previously established diabetic models. After 4wk of treatment, morphological alterations of Müller cells and protein expression of glutamine synthase(GS) and glial fibrillary acidic protein(GFAP)were analyzed.RESULTS:PEDFateither0.1μg/μLor0.2μg/μLsignificantly improved the structures of both nuclei and organelles of Müller cells compared to the BSS-treated group.Expression of GS was significantly higher in the 0.2 μg/μL PEDF group than that in the BSS group(P =0.012), but expression of GFAP was significantly lower in the 0.2 μg/μL PEDF group than that in the BSS group(P =0.000);however, there were no significant differences in expression of these proteins between the 0.1 μg/μL PEDF group and the BSS group(P =0.608, P =0.152). CONCLUSION: PEDF protects the morphological ultrastructure of Müller cells, improves the expression of glutamate synthase and prevents cell gliosis.

Filamentation of Metabolic Enzymes in Saccharomyces cerevisiae

Compartmentation via filamentation has recently emerged as a novel mechanism for metabolic regulation. In order to identify filamentforming metabolic enzymes systematically, we performed a genome-wide screening of all strains available from an open reading frameGFP collection in Saccharomyces cerevisiae. We discovered nine novel filament-forming proteins and also confirmed those identified previously. From the 4159 strains, we found 23 proteins, mostly metabolic enzymes, which are capable of forming filaments in vivo. In silico protein-protein interaction analysis suggests that these filament-forming proteins can be clustered into several groups, including translational initiation machinery and glucose and nitrogen metabolic pathways. Using glutamine-utilising enzymes as examples, we found that the culture conditions affect the occurrence and length of the metabolic filaments. Furthermore, we found that two CTP synthases（Ura7p and Ura8p） and two asparagine synthetases（Asn1p and Asn2p） form filaments both in the cytoplasm and in the nucleus.Live imaging analyses suggest that metabolic filaments undergo sub-diffusion. Taken together, our genome-wide screening identifies additional filament-forming proteins in S. cerevisiae and suggests that filamentation of metabolic enzymes is more general than currently appreciated.