Isolation and characterization of dinucleoside tetra- and tri-phosphates formed in the presence of lysyl-sRNA synthetase

Randerath, Kurt; Janeway, Claude M.; Stephenson, Mary L.; Zamecnik, Paul C.

doi:10.1016/0006-291x(66)90416-5

Cited by 136 publications

(45 citation statements)

References 12 publications

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“…LysRS has been known for 3 decades to be the main cellular source of Ap 4 A from both prokaryotes and eukaryotes (30,46,47). In our previous and present studies, we have revealed the biological significance of LysRS as an enzyme that participates in intracellular signaling.…”

Section: Discussionsupporting

confidence: 52%

Nonconventional Involvement of LysRS in the Molecular Mechanism of USF2 Transcriptional Activity in FcεRI-Activated Mast Cells

Lee¹,

Razin

2005

Molecular and Cellular Biology

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Reports of the biological multifunctional activity of various aminoacyl tRNA synthetases have recently accumulated in the literature. The primary function of these critical enzymes is to charge various tRNAs with their appropriate amino acids, thus producing the building blocks of protein synthesis. We have previously shown that lysyl tRNA synthetase (LysRS) associates with microphthalmia transcription factor (MITF) and regulates its activity by synthesis of Ap 4 A in mast cells. Here, we show for the first time that LysRS associates with another transcription factor, USF2, which unlike MITF, is ubiquitously expressed in eukaryotic cells. Using mast cells, we have found that USF2 is negatively regulated by Hint and Ap 4 A acts as a positive regulator of USF2 by a molecular mechanism similar to that described for MITF. Since USF2 plays a significant role in a variety of cellular functions, our finding suggests that LysRS and Ap 4 A may be involved in general regulation of gene transcription.

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Section: Discussionsupporting

confidence: 52%

Nonconventional Involvement of LysRS in the Molecular Mechanism of USF2 Transcriptional Activity in FcεRI-Activated Mast Cells

Lee¹,

Razin

2005

Molecular and Cellular Biology

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“…The binding activity of this protein is highly specific for Ap4A. Diadenosine 5',5"'-P',P4-tetraphosphate (Ap4A) was discovered in 1966 as a product of the back reaction of the amino acid activation step in protein synthesis (1,2). Some recent evidence implicates a possible participation of this dinucleotide in the process of DNA replication in eukaryotic cells.…”

mentioning

confidence: 99%

Resolution of the diadenosine 5',5"'-P1,P4-tetraphosphate binding subunit from a multiprotein form of HeLa cell DNA polymerase alpha.

Baril¹,

Bonin²,

Burstein³

et al. 1983

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

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A diadenosine 5',5"'-P1,P4-tetraphosphate (Ap4A) binding subunit has been resolved from a high molecular weight (640,000) multiprotein form of DNA polymerase alpha [deoxynucleoside triphosphate:DNA nucleotidyltransferase (DNA-directed), EC 2.7.7.7] from HeLa cells [DNA polymerase alpha 2 of Lamothe, P., Baril, B., Chi, A., Lee, L. & Baril, E. (1981) Proc. Natl. Acad. Sci. USA 78, 4723-4727]. The Ap4A binding activity copurifies with the DNA polymerizing activity during the course of purification. Hydrophobic chromatography on butylagarose resolves the Ap4A binding activity from the DNA polymerase. The Ap4A binding activity is protein in nature since the binding of Ap4A is abolished by treatment of the isolated binding activity with proteinase K but is insensitive to treatment with DNase or RNase. The molecular weight of the Ap4A binding protein, as determined by polyacrylamide gel electrophoresis under nondenaturing conditions or by NaDodSO4/polyacrylamide gel electrophoresis after photoaffinity labeling of the protein with [32P]Ap4A is 92,000 or 47,000. The binding activity of this protein is highly specific for Ap4A.

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“…The integrators used recognize, and quantitate such tangent peaks most easily when they are as narrow as in Figure 3. The observations on the shape of the ApaA peak at different pH values taken together with the fact that Ap4A is extremely stable at an alkaline pH (no hydrolysis in 0.24 M-LiOH at 30 ~ for one hour) (14), indicates that the present method of analysis is preferable, also because of its simplicity.…”

Section: Discussionmentioning

confidence: 85%

Determination of diadenosine tetraphosphate in biological material by high pressure liquid chromatography

Plesner¹,

Ottesen

1980

Carlsberg Res. Commun.

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A method for determination of diadenosine tetraphosphate, Ap4A, by high pressure liquid chromatography has been developed. The free nucleotides were extracted from biological material with I 0 % trichloroacetic acid, which subsequently was removed with n-trioctylamine dissolved in freon, the aquous phase containing the nucleotides was lyophilized. The lyophilized material was dissolved in 0. I M-NaHCO3, digested with alkaline phosphatase, injected into a strong anion exchange column without adjustment of pH and chromatographed isocratically with a KH2PO4-KCI buffer at pH 4.3. The enzymatic digestion of the cellular nucleoside mono-, di-, and triphosphates was followed and found to be completed in two hours, while Ap4A was observed not to be cleaved during this period. Amounts as small as 5-10 picomoles of Ap4A were determined quantitatively by the method described.

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Isolation and characterization of dinucleoside tetra- and tri-phosphates formed in the presence of lysyl-sRNA synthetase

Cited by 136 publications

References 12 publications

Nonconventional Involvement of LysRS in the Molecular Mechanism of USF2 Transcriptional Activity in FcεRI-Activated Mast Cells

Nonconventional Involvement of LysRS in the Molecular Mechanism of USF2 Transcriptional Activity in FcεRI-Activated Mast Cells

Resolution of the diadenosine 5',5"'-P1,P4-tetraphosphate binding subunit from a multiprotein form of HeLa cell DNA polymerase alpha.

Determination of diadenosine tetraphosphate in biological material by high pressure liquid chromatography

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