Binding of Spermine to tRNA<sup>Tyr</sup> Stabilizes the Conformation of the Anticodon Loop and Creates Strong Binding Sites for Divalent Cations

Nöthig-Laslo, Vesna; Živković, Tomislav P.; Kućan, Željko; Weygand-Đurašević, Ivana

doi:10.1111/j.1432-1033.1981.tb06332.x

Cited by 29 publications

(8 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This finding supports the notion that the above analogues cannot cause displacement of Mg 2ϩ from the ribosomal class (II) sites suggested by Weiss et al [36]. Also, an hypothetical displacement of Mg 2ϩ from AcPhe-tRNA sites does not seem possible, since Nothig-Laslo et al [42] have found that the parent compound, spermine, does not compete with divalent cations for common binding sites on tRNA.…”

Section: Discussionsupporting

confidence: 86%

Structure/function correlation of spermine‐analogue‐induced modulation of peptidyltransferase activity

Karahalios

Mamos

Karigiannis

et al. 1998

European Journal of Biochemistry

View full text Add to dashboard Cite

In a cell-free system derived from Escherichia coli,various analogues of spermine were used to study their effect on the binding of AcPhe-tRNA to poly (U)-programmed ribosomes and on the puromycin reaction carried out at 6 mM Mg 2ϩ (Ac, acetyl). In the absence of factors washable from ribosomes (FWR fraction), mono-acylated or di-acylated analogues of spermine stimulate the binding of AcPhe-tRNA to a lesser degree than spermine, in the order: N 1 -acetylspermine Ͼ N 1 ,N 12 -diacetylspermine Х N 1 ,N 12 -dipivaloylspermine. Also, the above analogues do not show any sparing effect on Mg 2ϩ requirements for AcPhe-tRNA binding to ribosomes, in contrast to spermine. The presence of FWR fraction during the binding or acetylation of the secondary amines of spermine moderates or abolishes the stimulatory effect.In addition, all analogues tested enhance the stability of the ternary complex AcPhe-tRNA-poly(U)-ribosome and the extent of AcPhe-puromycin synthesis, particularly in the absence of the FWR fraction. At the kinetic phase of AcPhe-puromycin synthesis, the analogues display both stimulatory and inhibitory effects, depending on the absence (partial noncompetitive inhibition) or the presence of the FWR fraction (nonessential activation in concert with partial noncompetitive inhibition). Detailed kinetic analysis shows that the analogues tested can mimic the behaviour of spermine, however, the potency to affect the peptidyltransferase activity depends on their degree of acylation, acyl-substituent size, charge distribution and on their chain flexibility.Keywords : acylated polyamine; protein synthesis; puromycin reaction; peptidyltransferase.Polyamines are organic polycations that are protonated at physiological pH and can potentially interact with a variety of cellular macromolecules including nucleic acids and proteins [1]. Substantial evidence exists suggesting a significant role of polyamines in regulating the translation process at several levels [1]. Due to its four positive charges, spermine is the most effective of the naturally occurring polyamines both in regulating the ribosomal functions and in decreasing the Mg 2ϩ requirements for protein synthesis. Tabor and Tabor [2] have reported that endogenous spermine does not exist in Escherichia coli. However, small amounts of spermine, about two orders of magnitude lower than those of spermidine, can be detected in E. coli by more sensitive methods [3]. Extensive biochemical studies have focused on the mechanism of spermine biological action in prokaryotic-cell-free systems, since this polyamine consists the parent compound of a large number of synthetic polyamine analogues with antiparasitic and antitumor activity [4Ϫ6].Evidently, spermine markedly stimulates the activity of several in vitro protein-synthesizing systems characterized by low Mg 2ϩ concentrations near those observed in vivo [7Ϫ10]. In a recent report [11], we have demonstrated that, at 6 mM Mg 2ϩ spermine exhibits a concentration-dependent allosteric biphasic

show abstract

Section: Discussionsupporting

confidence: 86%

Structure/function correlation of spermine‐analogue‐induced modulation of peptidyltransferase activity

Karahalios

Mamos

Karigiannis

et al. 1998

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…Other mechanisms of protein induction are also possible to be influenced by polyamines. Anticodon loop configuration of tRNA TM is claimed to be stabilized by the binding of spermidine [22]. DNA synthesis by type C reteroviral DNA polymerase was augmented 10 times by 0.5 mM spermine or by 2 mM spermidine [23].…”

Section: Discussionmentioning

confidence: 99%

Anti-inflammatory effect of polyamines in serotonin and carrageenan paw edemata—possible mechanism to increase vascular permeability inhibitory protein level which is regulated by glucocorticoids and superoxide radical

Òyanagui

1984

Agents and Actions

View full text Add to dashboard Cite

Serotonin paw edema of mice and carrageenan paw edema of rats were inhibited by subcutaneously or orally administered certain polyamines. They must be given at least 2 h before serotonin challenge to get inhibitions which were blocked by the concomitant injections of cycloheximide. Thirty percent inhibitory dose (ID30) of polyamines (s.c.) 3 h before serotonin (s.c.) were: spermidine (8 mg/kg), spermine 28 mg/kg) and putrescine (55 mg/kg). Agmatine, cadaverine, ornithine, citrulline, lysine and arginine were not inhibitory even at 200 mg/kg. Three inhibitory polyamines were effective by oral administration but were not inhibitory by local administration into the paws. Intravenous injections of spermidine also required 2 h of lag period for inhibitions. Serotonin edema was inhibited by dexamethasone (1 mg/kg), prednisolone (1 mg/kg) or by superoxide dismutase (SOD, 5 mg/kg) in lag period requiring manner (s.c. and i.v.). High dose of cyclo-oxygenase inhibitors indomethacin and diclofenac sodium, lipo-oxygenase inhibitor BW755C (30 mg/kg s.c., respectively) and phospholipase A2 inhibitor quinacrine (100 mg/kg s.c.) failed to inhibit serotonin edema, suggesting that arachidonate metabolites are not participating in this model. ID30 of polyamines which were administered (s.c. and oral) to rats 3 h before carrageenan and determined at 3 h by paw weight were: spermidine (28 and 100 mg/kg), spermine (18 and 90 mg/kg) and putrescine (both greater than 200 mg/kg). Adrenalectomized rats responded to polyamines just as normal rats. Local vascular permeability, irritancy and acute toxicity were also tested in mice. Polyamines were proved to be glucocorticoid-type anti-inflammatory drugs. Polyamines may be mediators of glucocorticoids for the synthesis of the postulated vascular permeability inhibitory protein (called as 'vasoregulin' for convenience). Anti-inflammatory effect of glucocorticoid is recently explained by its capacity to induce phospholipase A2 inhibitory protein(s) (macrocortin or lipomodulin). However, this hypothesis has not yet been proved by in vivo experiment and our data suggest that there is induction by glucocorticoid of another kind of protein which does not inhibit phospholipase A2 activity.

show abstract

“…There have been a number of studies on the effects of polyamines on various steps in protein synthesis in vitro, including initiation (19,20) and translation (21), on tRNA structure (22)(23)(24)(25), on tRNA aminoacylation (26), and on the fidelity of translation (27)(28)(29). (For additional references see the articles cited in refs.…”

Section: Discussionmentioning

confidence: 99%

Polyamine requirement for efficient translation of amber codons in vivo.

Tabor¹,

Tabor²

1982

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Multiplication of several amber mutants of bacteriophage T7 was decreased in two polyamine-deficient mutants of Escherichia coli K-12 carrying amber suppressors, relative to the multiplication ofwild type bacteriophage T7 in the same hosts. In contrast the same T7 amber bacteriophages multiplied well in these strains when supplemented with polyamines. The requirement for polyamines for optimal translation of amber codons in vivo was confirmed by showing that infection of polyamine-depleted E. coli with bacteriophage T7 carrying an amber mutation in gene I resulted in an increased accumulation ofthe amber fragment of the gene 1 protein and a decreased accumulation of the full-length gene 1 protein compared with infection of an aminesupplemented culture. These results indicate that one important function of polyamines in vivo is concerned with protein translation and the protein-synthesizing ribosomal complex.Although many effects ofputrescine and spermidine have been observed in in vitro systems (for references, see refs. 1-3), we still do not know the biological role of these amines in vivo. For this reason we have developed mutants of Escherichia coli that have deletions in the genes for the biosynthetic enzymes for these amines and hence contain no amines when grown in an amine-free medium (4, 5). The organisms that we first described could grow indefinitely in the absence of amines but did so at a lowered growth rate.One of the postulated roles for polyamines is an effect on some aspect of protein biosynthesis, and we decided to see if a role in protein synthesis could be demonstrated in amine-deficient mutants in vivo. In previous experiments (6), we showed that transduction of certain rpsL (strA)* alleles into an aminerequiring strain converted this organism from one with a partial requirement for amines for growth into one with an absolute requirement. Because rpsL mutations are known to cause defects in the S12 ribosomal protein and to affect the fidelity of translation in vitro and in vivo (8-10), our studies suggested that polyamines might act by affecting the stability and conformation of the protein-synthesizing ribosomal complex.The experiments in the present paper examined the effect of polyamines on translation from a different point of view: namely, does polyamine deprivation affect the translation of amber codons in vivo? We have shown that, in E. coli, polyamine deficiency causes a marked decrease in the multiplication of some amber mutants of bacteriophage T7, compared to the multiplication ofwild type bacteriophage T7, even though these bacterial strains contain a suppressor. However, when these strains are grown in the presence of polyamines, they are suitable hosts for the amber mutants of bacteriophage T7 as well as the wild type. These results indicate that polyamines are required for the optimal translation of an amber codon in a strain carrying a suppressor and support the hypothesis that an important function of the polyamines is concerned with the protein-synthesizing ribosom...

show abstract

Binding of Spermine to tRNA^Tyr Stabilizes the Conformation of the Anticodon Loop and Creates Strong Binding Sites for Divalent Cations

Cited by 29 publications

References 16 publications

Structure/function correlation of spermine‐analogue‐induced modulation of peptidyltransferase activity

Structure/function correlation of spermine‐analogue‐induced modulation of peptidyltransferase activity

Anti-inflammatory effect of polyamines in serotonin and carrageenan paw edemata—possible mechanism to increase vascular permeability inhibitory protein level which is regulated by glucocorticoids and superoxide radical

Polyamine requirement for efficient translation of amber codons in vivo.

Contact Info

Product

Resources

About

Binding of Spermine to tRNATyr Stabilizes the Conformation of the Anticodon Loop and Creates Strong Binding Sites for Divalent Cations

Cited by 29 publications

References 16 publications

Structure/function correlation of spermine‐analogue‐induced modulation of peptidyltransferase activity

Structure/function correlation of spermine‐analogue‐induced modulation of peptidyltransferase activity

Anti-inflammatory effect of polyamines in serotonin and carrageenan paw edemata—possible mechanism to increase vascular permeability inhibitory protein level which is regulated by glucocorticoids and superoxide radical

Polyamine requirement for efficient translation of amber codons in vivo.

Contact Info

Product

Resources

About

Binding of Spermine to tRNA^Tyr Stabilizes the Conformation of the Anticodon Loop and Creates Strong Binding Sites for Divalent Cations