Richaud C, Zabulon G, Joder A, Thomas J C
Unité Mixte de Recherche 8543, Centre National de la Recherche Scientifique, "Photorégulation et Dynamique des Membranes Végétales," Ecole Normale Supérieure, 75230 Paris cedex 05, France.
J Bacteriol. 2001 May;183(10):2989-94. doi: 10.1128/JB.183.10.2989-2994.2001.
Nitrogen (N) limitation in cyanobacteria is well documented: a reduced growth rate is observed, accompanied by a cessation of phycobiliprotein synthesis and an ordered degradation of phycobilisomes (PBS). This leads to a dramatic bleaching phenomenon known as chlorosis. In Synechococcus strain PCC 7942, bleaching due to PBS degradation is also observed under sulfur (S) or phosphorus (P) limitation, and all three are under the control of the nblA gene product, a 59-amino-acid polypeptide which is overexpressed under N, S, and P starvation (J. L. Collier, and A. R. Grossman, EMBO J. 13:1039-1047, 1994). Cyanobase sequence data for Synechocystis strain PCC 6803 indicate the presence of two tandem open reading frames (sll0452 and sll0453) homologous to nblA. We cloned the two genes, identified a unique 5' mRNA end suggestive of a single transcription start site, and studied nblA expression under conditions of N or S starvation by Northern hybridization: transcripts were detected only under N starvation (no signal is detected in replete medium or with S starvation), whether nblA1 or nblA2 was used as a probe. Mutations in nblA1 and nblA2 were constructed by insertion of a kanamycin cassette; both mutations were nonbleaching under N starvation. Synechocystis strain PCC 6803 does not bleach under S starvation, consistent with the absence of nblA induction in these conditions. These results were confirmed by analysis of the PBS components: sequential degradation of phycocyanin and associated linkers was observed only under conditions of N starvation. This indicates differences between Synechocystis strain PCC 6803 and Synechococcus strain PCC 7942 in their regulatory and signaling pathways leading to N- and S-starved phenotypes.
蓝藻中氮(N)限制的现象已有充分记录:观察到生长速率降低,同时伴随着藻胆蛋白合成的停止以及藻胆体(PBS)的有序降解。这会导致一种显著的白化现象,即黄化。在聚球藻菌株PCC 7942中,在硫(S)或磷(P)限制条件下也会观察到由于PBS降解导致的白化现象,并且这三种情况都受nblA基因产物的控制,该基因产物是一种59个氨基酸的多肽,在氮、硫和磷饥饿条件下会过度表达(J. L. 科利尔和A. R. 格罗斯曼,《欧洲分子生物学组织杂志》13:1039 - 1047, 1994)。集胞藻菌株PCC 6803的蓝藻碱基序列数据表明存在两个与nblA同源的串联开放阅读框(sll0452和sll0453)。我们克隆了这两个基因,确定了一个独特的5' mRNA末端,提示存在一个单一的转录起始位点,并通过Northern杂交研究了在氮或硫饥饿条件下nblA的表达:无论使用nblA1还是nblA2作为探针,转录本仅在氮饥饿条件下被检测到(在营养充足的培养基中或硫饥饿条件下未检测到信号)。通过插入卡那霉素盒构建了nblA1和nblA2的突变体;在氮饥饿条件下,这两个突变体均不发生白化。集胞藻菌株PCC 6803在硫饥饿条件下不会白化,这与在这些条件下未诱导nblA一致。通过对PBS成分的分析证实了这些结果:仅在氮饥饿条件下观察到藻蓝蛋白和相关连接子的顺序降解。这表明集胞藻菌株PCC 6803和聚球藻菌株PCC 7942在导致氮和硫饥饿表型的调控和信号传导途径上存在差异。