Effect of Acetylpolyamines on In Vitro Protein Synthesis and on the Growth of a Polyamine-Requiring Mutant of Escherichia coli

Kakegawa, Tomohito; Guo, Yan; Chiba, Yuko; Miyazaki, Toshiko; Nakamura, Mako; Hirose, Sachiko; Canellakis, Zoe N.; Igarashi, Kazuei

doi:10.1093/oxfordjournals.jbchem.a123431

Cited by 26 publications

(32 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Our evidence suggests that acetylation occurs at low temperature to prevent excess spermidine from inhibiting the binding of Mg 2ϩ to the ribosome. Consistent with this is the finding that acetylpolyamines cannot substitute for polyamines in binding to RNA (18). Although the acetylpolyamine is still cationic, it has been suggested that the acetyl group may present a steric hindrance to the binding of nucleic acids.…”

Section: Discussionmentioning

confidence: 64%

“…An effect of polyamines in in vitro translation is the lowering of the Mg 2ϩ requirement (18). Since spermidine binds to sites on the ribosome similar to those bound by Mg 2ϩ , it can replace ribosome-bound Mg 2ϩ (5,28).…”

Section: Resultsmentioning

confidence: 99%

“…Furthermore, the excess spermidine causes decreased protein synthesis and cell viability during the stationary phase of growth (9). Acetylation serves to convert the polyamine to a physiologically inert form; acetylpolyamines cannot substitute for polyamines in RNA binding, in the enhancement of growth of an E. coli polyamine-deficient mutant, or in the stimulation of in vitro translation (18). Neither spermidine-deacetylating activity nor polyamine oxidase activity has been detected in E. coli, suggesting that the N-acetylspermidine is either excreted or kept in the inert acetylated form (14,24).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Spermidine Acetyltransferase Is Required To Prevent Spermidine Toxicity at Low Temperatures in Escherichia coli

Limsuwun

Jones

2000

J Bacteriol

View full text Add to dashboard Cite

Polyamines are required for optimal growth in most cells; however, polyamine accumulation leads to inhibition of cellular growth. To reduce intracellular polyamine levels, spermidine is monoacetylated in both prokaryotes and eukaryotes. In Escherichia coli, the speG gene encodes the spermidine acetyltransferase, which transfers the acetyl group to either the N-1 or N-8 position. In addition to polyamine accumulation, stress conditions, such as cold shock, cause an increase in the level of spermidine acetylation, suggesting an adaptive role for reduced polyamine levels under stressful growth conditions. The effect of spermidine accumulation on the growth of E. coli at low temperature was examined using a speG mutant. At 37°C, growth of the speG mutant was normal in the presence of 0.5 or 1 mM spermidine. However, following a shift to 7°C, the addition of 0.5 or 1 mM spermidine resulted in inhibition of cellular growth or cell lysis, respectively. Furthermore, at 7°C, spermidine accumulation resulted in a decrease in total protein synthesis accompanied by an increase in the synthesis of the major cold shock proteins CspA, CspB, and CspG. However, the addition of 50 mM Mg Polyamines, such as spermidine and putrescine, are present in virtually all cells (25, 33). These polycations have pleiotropic properties, which can influence several cellular processes. They can bind to nucleic acids, stabilize membranes, and stimulate the activity of several enzymes, such as RNA polymerase (1,31,34). Found in the ribosomal fraction, polyamines enhance the synthesis of several proteins and stimulate the in vivo assembly of the Escherichia coli 30S ribosomal subunit (7,13,21,35).Although intracellular polyamine is required for optimal growth, polyamine accumulation can lead to inhibition of cellular growth (11,25,26,33). The addition of spermidine to cell cultures of mouse FM3A cells results in a decrease in cell growth accompanied by inhibition of protein synthesis (11). To prevent polyamine toxicity in eukaryotes, polyamine catabolism is initiated by the monoacetylation of spermidine and spermine catalyzed by spermidine/spermine N 1 -acetyltransferase (SSAT) (4). The acetylpolyamines are then either further oxidized by polyamine oxidase or excreted from the cell.In E. coli, the speG gene encodes spermidine acetyltransferase (SAT), which transfers the acetyl group to either the N-1 or N-8 position of the polyamine (2,8,20). SAT is required to reduce the spermidine level, since the addition of exogenous spermidine to a speG mutant results in intracellular accumulation of spermidine (9). Furthermore, the excess spermidine causes decreased protein synthesis and cell viability during the stationary phase of growth (9). Acetylation serves to convert the polyamine to a physiologically inert form; acetylpolyamines cannot substitute for polyamines in RNA binding, in the enhancement of growth of an E. coli polyamine-deficient mutant, or in the stimulation of in vitro translation (18). Neither spermidine-deacetylating activity nor polyamin...

show abstract

Section: Discussionmentioning

confidence: 64%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Spermidine Acetyltransferase Is Required To Prevent Spermidine Toxicity at Low Temperatures in Escherichia coli

Limsuwun

Jones

2000

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…Abnormal accumulation of polyamine, however, can lead to inhibition of cell growth and protein synthesis (29); therefore, polyamine concentration should be regulated strictly and promptly to maintain optimum cell growth. It was reported previously that N-acetylpolyamine had less effect on growth and protein synthesis than non-acetylated polyamine (30), and a pathway to synthesize N-acetylpolyamine to decrease the toxicity of excess polyamine exists in both prokaryotes and eukaryotes (6,31). Because the ␥-glutamylation of putrescine by PuuA is similar to the acetylation of polyamines, it is predicted that PuuA also plays a role in detoxifying excess polyamine.…”

Section: Discussionmentioning

confidence: 99%

γ-Glutamylputrescine Synthetase in the Putrescine Utilization Pathway of Escherichia coli K-12

Kurihara

Oda

Tsuboi

et al. 2008

Journal of Biological Chemistry

110

View full text Add to dashboard Cite

Glutamate-putrescine ligase (␥-glutamylputrescine synthetase, PuuA, EC 6.3.1.11) catalyzes the ␥-glutamylation of putrescine, the first step in a novel putrescine utilization pathway involving ␥-glutamylated intermediates, the Puu pathway, in Escherichia coli. In this report, the character and physiological importance of PuuA are described. Purified non-tagged PuuA catalyzed the ATP-dependent ␥-glutamylation of putrescine. The K m values for glutamate, ATP, and putrescine are 2.07, 2.35, and 44.6 mM, respectively. There are two putrescine utilization pathways in E. coli: the Puu pathway and the pathway without ␥-glutamylation. Gene deletion experiments of puuA, however, indicated that the Puu pathway was more critical in utilizing putrescine as a sole carbon or nitrogen source. The transcription of puuA was induced by putrescine and in a puuR-deleted strain. The amino acid sequences of PuuA and glutamine synthetase (GS) show high similarity. The molecular weights of the monomers of the two enzymes are quite similar, and PuuA exists as a dodecamer as does GS. Moreover the two amino acid residues of E. coli GS that are important for the metal-catalyzed oxidation of the enzyme molecule involved in protein turnover are conserved in PuuA, and it was experimentally shown that the corresponding amino acid residues in PuuA were involved in the metal-catalyzed oxidation similarly to GS. It is suggested that the intracellular concentration of putrescine is optimized by PuuA transcriptionally and posttranslationally and that excess putrescine is converted to a nutrient source by the Puu pathway.

show abstract

“…Unlike spermidine, N-acetylspermidine is physiologically inert and does not bind to RNA or stimulate in vitro translation. Therefore, active conversion of spermidine to N-acetylspermidine by spermidine acetyltransferase would be an effective mechanism to reduce the toxicity associated with high levels of spermidine (Kakegawa et al, 1991). Although speculative, it is possible that cell density-dependent changes in intracellular spermidine levels, which are linked to methionine utilization and the production of S-adenosylmethionine (SAM), function as a metabolic sensor for B. pseudomallei to regulate gene expression and protein synthesis in a growth phase-dependent manner.…”

Section: Discussionmentioning

confidence: 99%

Growth-related changes in intracellular spermidine and its effect on efflux pump expression and quorum sensing in Burkholderia pseudomallei

Chan

Chua

2010

Microbiology

View full text Add to dashboard Cite

The Burkholderia pseudomallei BpeAB-OprB resistance-nodulation-division (RND) family pump effluxes aminoglycoside and macrolide antibiotics as well as acylhomoserine lactones (AHLs) involved in quorum sensing. Expression of bpeA-lacZ was cell density-dependent and was inducible in the presence of these compounds. Intracellular levels of spermidine and N-acetylspermidine increased with cell density in wild-type B. pseudomallei KHW, but were always lower in the bpeAB pump mutant at all growth phases. The significance of changes in intracellular spermidine on efflux pump expression was demonstrated by the disruption of the binding of the BpeR repressor protein to the bpeABoprB regulatory region in vitro in the presence of increasing spermidine concentrations. This was supported by dose-dependent activation of bpeA-lacZ transcription in vivo in the presence of exogenous spermidine and N-acetylspermidine, thus implicating the involvement of the BpeAB-OprB pump in spermidine homeostasis in B. pseudomallei. Consequently, inhibition of intracellular spermidine synthesis reduced the efflux of AHLs by BpeAB-OprB. Other potential therapeutic applications of spermidine synthase inhibitors include the reduction of swimming motility and biofilm formation by B. pseudomallei.

show abstract

Effect of Acetylpolyamines on In Vitro Protein Synthesis and on the Growth of a Polyamine-Requiring Mutant of Escherichia coli

Cited by 26 publications

References 0 publications

Spermidine Acetyltransferase Is Required To Prevent Spermidine Toxicity at Low Temperatures in Escherichia coli

Spermidine Acetyltransferase Is Required To Prevent Spermidine Toxicity at Low Temperatures in Escherichia coli

γ-Glutamylputrescine Synthetase in the Putrescine Utilization Pathway of Escherichia coli K-12

Growth-related changes in intracellular spermidine and its effect on efflux pump expression and quorum sensing in Burkholderia pseudomallei

Contact Info

Product

Resources

About