The AGUAAA motif in cspA1/A2 mRNA is important for adaptation of Yersinia enterocolitica to grow at low temperature
The AGUAAA motif in cspA1/A2 mRNA is important for adaptation of Yersinia enterocolitica to grow at low temperature
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DOI：
10.1046/j.1365-2958.2003.03795.x
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发表时间：
2003-12-01
影响因子：
3.6
通讯作者：
Scherer, S
中科院分区：
文献类型：
--
作者：
Neuhaus, K;Anastasov, N;Scherer, S
耐冷性小肠结肠炎耶尔森氏菌在30 ℃至10 ℃的冷休克后的适应导致主要冷休克蛋白（CSP）双顺反子基因cspA 1/A2的转录增加高达300倍。使用与CSP mRNA的不同区域特异性杂交的四种探针对cspA 1/A2进行的北方印迹分析揭示了许多cspA 1/A2转录物的出现，这些转录物小于原始转录物，并且在驯化期结束时短暂可见。引物延伸和RNA保护实验表明，这些较小的mRNA具有位于mRNA内五个不同位置的相同核心序列（5 '-AGUAAA-3'）中的5'末端，表明CSP mRNA转录物的优先切割。对于含有两个这样的核心序列的大肠杆菌的cspB获得了类似的结果。此外，该基序存在于多种革兰氏阴性和革兰氏阳性细菌的主要CSP基因中。因此，我们把这一序列称为冷冲击切割盒（CSC盒）。在Y.在小肠结肠炎中，该构建体的mRNA在CSC盒内被切割，并且该CSC盒从AGUAAA到AGUCCC的变化显著降低了突变lacZ基因的切割。在Y中突变所有CSC框。在小肠结肠炎中，质粒结合的cspA 1/A2显著增加了冷休克后再生长发生之前的滞后时间。基于这些结果，我们认为CSC盒的作用与低温适应后cspA mRNA的下调有关。
Acclimatization of the psychrotolerant Yersinia enterocolitica after a cold shock from 30degreesC to 10degreesC causes transcription of the major cold shock protein (CSP) bicistronic gene cspA1/A2 to increase by up to 300-fold. Northern blot analysis of cspA1/A2 using four probes that hybridize specifically to different regions of CSP mRNA revealed the appearance of a number of cspA1/A2 transcripts that are smaller than the original transcript and transiently visible at the end of the acclimation period. Primer extension and RNA protection experiments demonstrated that these smaller mRNAs have 5' ends located in the same core sequence (5'-AGUAAA-3') at five different places within the mRNA, indicating preferential cleavage of the CSP mRNA transcripts. A similar result was obtained for cspB of Escherichia coli, containing two such core sequences. Furthermore, this motif is present in the major CSP genes of a variety of Gram-negative and Gram-positive bacteria. We have therefore termed this sequence cold shock cut box (CSC-box). After inserting a CSC-box into a plasmid-bound lacZ gene in Y. enterocolitica, the mRNA of this construct was cleaved within the CSC-box, and a change in this CSC-box from AGUAAA to AGUCCC dramatically reduced cleavage of the mutated lacZ gene. Mutating all CSC-boxes in Y. enterocolitica of a plasmid bound cspA1/A2 dramatically increases the lag time after a cold shock before re-growth occurs. Based on these results, we suggest that the role of the CSC-box is related to downregulation of cspA mRNA after acclimation to low temperature.