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Properties of putrescine uptake by PotFGHI and PuuP and their physiological significance in Escherichia coli

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Abstract

Properties of putrescine uptake by PotFGHI and PuuP and their physiological significance were studied using a polyamine biosynthesis and uptake deficient Escherichia coli KK3131 transformed with pACYC184 containing potFGHI or puuP. Putrescine uptake activity of E. coli KK3131 transformed with pACYC184-PotFGHI was higher than that of E. coli 3131 transformed with pACYC-PuuP when cells were cultured in the absence of putrescine. Putrescine uptake by PotFGHI was both ATP and membrane potential dependent, while that by PuuP was membrane potential dependent. Feedback inhibition by polyamines occurred at the PotFGHI uptake system but not at the PuuP uptake system. Expression of PuuP was reduced in the presence of PuuR, a negative regulator for PuuP, and expression of PuuR was positively regulated by glucose, which reduces the level of cAMP. The complex of cAMP and CRP (cAMP receptor protein) inhibited the expression of PuuR in the absence of glucose. Thus, the growth rate of E. coli KK3131 in the presence of both 0.4 % (22.2 mM) glucose and 10 mM putrescine was in the order of cells transformed with pACYC-PotFGHI > pACYC-PuuP > pACYC-PuuP + PuuR, which was parallel with the polyamine content in cells. The results indicate that PotFGHI is necessary for rapid cell growth in the presence of glucose as an energy source. When glucose in medium was depleted, however, PuuP was absolutely necessary for cell growth in the presence of putrescine, because accumulation of putrescine to a high level by PuuP was necessary for utilization of putrescine as an energy source.

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Abbreviations

CadB:

Cadaverine-lysine antiporter

CCCP:

Carbonyl cyanide m-chlorophenylhydrazone

CRP:

cAMP receptor protein

PotE:

Putrescine-ornithine antiporter

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Acknowledgments

We thank Drs. A. J. Michael and K. Williams for their help in preparing the manuscript. We also thank Dr. A. Ishihama for providing antibodies against CRP and RNA polymerase σ70 subunit. This work was supported by a Gran-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, Culture, and Technology, Japan. S. D. Saroj was supported by the Tokyo Biochemical Research Foundation Research Fellowship.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Faculty of Pharmacy, Chiba Institute of Science, Shiomi-cho, Choshi, Chiba, Japan

    Yusuke Terui, Akihiko Sakamoto, Taketo Yoshida & Keiko Kashiwagi

  2. Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1, Inohana, Chuo-ku, Chiba, 260-8675, Japan

    Sunil D. Saroj, Kyohei Higashi, Toshihiko Toida & Kazuei Igarashi

  3. Graduate School of Science and Technology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto, Japan

    Shin Kurihara & Hideyuki Suzuki

  4. Amine Pharma Research Institute, Innovation Plaza at Chiba University, Inohana, Chuo-ku, Chiba, Japan

    Kazuei Igarashi

Authors
  1. Yusuke Terui
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  2. Sunil D. Saroj
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  3. Akihiko Sakamoto
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  4. Taketo Yoshida
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  5. Kyohei Higashi
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  6. Shin Kurihara
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  7. Hideyuki Suzuki
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  8. Toshihiko Toida
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  9. Keiko Kashiwagi
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  10. Kazuei Igarashi
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Correspondence to Kazuei Igarashi.

Additional information

Y. Terui and S. D. Saroj contributed equally to this work.

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Terui, Y., Saroj, S.D., Sakamoto, A. et al. Properties of putrescine uptake by PotFGHI and PuuP and their physiological significance in Escherichia coli . Amino Acids 46, 661–670 (2014). https://doi.org/10.1007/s00726-013-1517-x

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  • DOI: https://doi.org/10.1007/s00726-013-1517-x

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