Zheng Zhenggao, Omairi-Nasser Amin, Li Xiying, Dong Chunxia, Lin Yan, Haselkorn Robert, Zhao Jindong
State Key Laboratory of Protein and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, People's Republic of China.
Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL 60637.
Proc Natl Acad Sci U S A. 2017 Feb 21;114(8):E1405-E1412. doi: 10.1073/pnas.1621424114. Epub 2017 Feb 3.
Channels that cross cell walls and connect the cytoplasm of neighboring cells in multicellular cyanobacteria are pivotal for intercellular communication. We find that the product of the gene of the filamentous cyanobacterium sp. PCC 7120 is required for proper channel formation. encodes an amidase that hydrolyses purified peptidoglycans. An All1140-GFP fusion protein is located at the Z-ring in the periplasmic space during most of the cell cycle. An -null mutant (M40) was unable to grow diazotrophically, and no mature heterocysts were observed in the absence of combined nitrogen. Expression of two key genes, and , was studied in M40 using GFP as a reporter. Upon nitrogen step-down, the patterned distribution of green fluorescent cells in filaments seen in the wild type were not observed in mutant M40. Intercellular communication in M40 was studied by measuring fluorescence recovery after photobleaching (FRAP). Movement of calcein (622 Da) was aborted in M40, suggesting that the channels connecting the cytoplasm of neighboring cells are impaired in the mutant. The channels were examined with electron tomography; their diameters were nearly identical, 12.7 nm for the wild type and 12.4 nm for M40, suggesting that AmiC3 is not required for channel formation. However, when the cell wall sacculi isolated by boiling were examined by EM, the average sizes of the channels of the wild type and M40 were 20 nm and 12 nm, respectively, suggesting that the channel walls of the wild type are expandable and that this expandability requires AmiC3.
在多细胞蓝细菌中,穿过细胞壁并连接相邻细胞细胞质的通道对于细胞间通讯至关重要。我们发现丝状蓝细菌集胞藻属PCC 7120的基因产物对于正确的通道形成是必需的。该基因编码一种可水解纯化肽聚糖的酰胺酶。在细胞周期的大部分时间里,一种All1140 - GFP融合蛋白位于周质空间的Z环处。一个该基因缺失的突变体(M40)无法进行固氮生长,并且在没有化合态氮的情况下未观察到成熟的异形胞。使用GFP作为报告基因,在M40中研究了两个关键基因的表达。在氮源减少时,突变体M40中未观察到野生型丝状藻丝中绿色荧光细胞的模式分布。通过测量光漂白后的荧光恢复(FRAP)研究了M40中的细胞间通讯。在M40中,钙黄绿素(622 Da)的移动受阻,这表明连接相邻细胞细胞质的通道在该突变体中受损。通过电子断层扫描检查通道;它们的直径几乎相同,野生型为12.7 nm,M40为12.4 nm,这表明通道形成不需要AmiC3。然而,当通过电子显微镜检查经煮沸分离的细胞壁囊泡时,野生型和M40通道的平均大小分别为20 nm和12 nm,这表明野生型的通道壁是可扩展的,并且这种可扩展性需要AmiC3。