Sir2B, A Sirtuin Family Protein, Involves in Cell Adhesion Molecule Expression During Early Dictyostelium Development Upon Starvation

H. Taniura, Shuhei Soeda, Ai Fujii, Mio Morioka, Minori Nakamura, Yui Sano
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Abstract

Sirtuin involves in cellular processes to adapt to starvation in the endocrine and metabolic systems. We examined Sir2B function during early Dictyostelium development upon starvation. GFP-tagged Sir2B was expressed and the immunoreactivity was detected a punctate pattern near cell surface. Sir2B mRNA was expressed in vegetative cells and peaked at 2 h after starvation. Ectopic expression of Sir2B enhanced aggregate formation with increased expressions of cell adhesion molecules such as cadA, csaA, TgrB1, and TgrC1 at 8h compared to those in control cells. Sir2B null-mutant cells (Sir2B KO cells) were generated and they formed aggregates smaller than those of wild-type cells after 24 h. Because cell-cell adhesion affects aggregation size, cell cohesion assay of Sir2B KO cells showed cell-cell adhesion impairment at 8 and 10 h during development, and the expressions of four cell adhesion molecules were reduced in Sir2B KO cells compared to wild-type cells until 8 h after starvation. When RNAi-mediated cadA or csaA knockdown cells (cadA KD or csaA KD cells) were generated, csaA KD cells formed smaller aggregates and affected the expressions of other cell adhesion molecules to reduce, but cadA KD cells did not show to affect aggregation size and cell adhesion molecules expressions apparently. These results suggest that Sir2B is involved in csaA expression and affects other cell adhesion molecules during early development upon starvation.
Sir2B,一种Sirtuin家族蛋白,参与饥饿时早期盘基骨菌发育过程中的细胞粘附分子表达
Sirtuin参与细胞在内分泌和代谢系统中适应饥饿的过程。我们检测了Sir2B在盘基骨菌饥饿发育早期的功能。表达gfp标记的Sir2B,并在细胞表面附近以点状模式检测免疫反应性。Sir2B mRNA在营养细胞中表达,并在饥饿后2 h达到峰值。与对照细胞相比,Sir2B的异位表达增强了聚集体的形成,并在8h时增加了细胞粘附分子如cadA、csaA、TgrB1和TgrC1的表达。产生Sir2B零突变细胞(Sir2B KO细胞),在24 h后形成比野生型细胞更小的聚集体。由于细胞间粘附影响聚集大小,Sir2B KO细胞的细胞内聚实验显示,在发育过程中的8和10 h, Sir2B KO细胞的细胞间粘附受损,直到饥饿后8 h, Sir2B KO细胞中4种细胞粘附分子的表达比野生型细胞减少。当产生rnai介导的cadA或csaA敲低细胞(cadA KD或csaA KD细胞)时,csaA KD细胞形成较小的聚集体,并影响其他细胞粘附分子的表达减少,但cadA KD细胞对聚集体大小和细胞粘附分子表达没有明显影响。这些结果表明Sir2B参与了csaA的表达,并在饥饿的早期发育过程中影响其他细胞粘附分子。
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