Yoshida Taketo, Sakamoto Akihiko, Terui Yusuke, Takao Koichi, Sugita Yoshiaki, Yamamoto Kaneyoshi, Ishihama Akira, Igarashi Kazuei, Kashiwagi Keiko
Faculty of Pharmacy, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba, 288-0025, Japan.
Laboratory of Bioorganic Chemistry, Department of Pharmaceutical Technology, Josai University, 1-1 Keyaki-dai, Sakado, Saitama, 350-0295, Japan.
PLoS One. 2016 Jul 19;11(7):e0159494. doi: 10.1371/journal.pone.0159494. eCollection 2016.
The effects of spermidine analogues [norspermidine (NSPD, 33), spermidine (SPD, 34), homospermidine (HSPD, 44) and aminopropylcadaverine (APCAD, 35)] on cell growth were studied using Escherichia coli polyamine-requiring mutant MA261. Cell growth was compared at 32°C, 37°C, and 42°C. All four analogues were taken up mainly by the PotABCD spermidine-preferential uptake system. The degree of stimulation of cell growth at 32°C and 37°C was NSPD ≥ SPD ≥ HSPD > APCAD, and SPD ≥ HSPD ≥ NSPD > APCAD, respectively. However, at 42°C, it was HSPD » SPD > NSPD > APCAD. One reason for this is HSPD was taken up effectively compared with other triamines. In addition, since natural polyamines (triamines and teteraamines) interact mainly with RNA, and the structure of RNA is more flexible at higher temperatures, HSPD probably stabilized RNA more tightly at 42°C. We have thus far found that 20 kinds of protein syntheses are stimulated by polyamines at the translational level. Among them, synthesis of OppA, RpoE and StpA was more strongly stimulated by HSPD at 42°C than at 37°C. Stabilization of the initiation region of oppA and rpoE mRNA was tighter by HSPD at 42°C than 37°C determined by circular dichroism (CD). The degree of polyamine stimulation of OppA, RpoE and StpA synthesis by NSPD, SPD and APCAD was smaller than that by HSPD at 42°C. Thus, the degree of stimulation of cell growth by spermidine analogues at the different temperatures is dependent on the stimulation of protein synthesis by some components of the polyamine modulon.
使用大肠杆菌多胺需求突变体MA261研究了亚精胺类似物[ norspermidine (NSPD, 33)、亚精胺(SPD, 34)、高亚精胺(HSPD, 44)和氨丙基尸胺(APCAD, 35)]对细胞生长的影响。比较了在32°C、37°C和42°C下的细胞生长情况。所有四种类似物主要通过PotABCD亚精胺优先摄取系统被摄取。在32°C和37°C时,细胞生长的刺激程度分别为NSPD≥SPD≥HSPD > APCAD和SPD≥HSPD≥NSPD > APCAD。然而,在42°C时,情况为HSPD » SPD > NSPD > APCAD。原因之一是与其他三胺相比,HSPD被有效摄取。此外,由于天然多胺(三胺和四胺)主要与RNA相互作用,并且RNA的结构在较高温度下更具柔性,HSPD可能在42°C时更紧密地稳定了RNA。到目前为止,我们发现有20种蛋白质合成在翻译水平上受到多胺的刺激。其中,OppA、RpoE和StpA的合成在42°C时比在37°C时受到HSPD的刺激更强。通过圆二色性(CD)测定,在42°C时,HSPD对oppA和rpoE mRNA起始区域的稳定作用比在37°C时更紧密。在42°C时,NSPD、SPD和APCAD对OppA、RpoE和StpA合成的多胺刺激程度小于HSPD。因此,亚精胺类似物在不同温度下对细胞生长的刺激程度取决于多胺调节子某些成分对蛋白质合成的刺激作用。