Effects of metals on the binding of protein factors to the mouse 2',5'- oligoadenylate synthetase ME-12 gene regulatory region

Cong Yan, I. Tamm

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Abstract

We have previously demonstrated that using mouse 2',5'-oligoadenylate synthetase (2-5A synthetase) gene (ME-12) regulatory region AB as a probe, at least six complexes (C1-C6) can be detected by the mobility shift electrophoresis assay with extracts of murine BALB/c-3T3 cells. The formation of these complexes is greatly influenced by different buffer conditions and the presence of metals. C1 and C4 signals are stronger in Tris-HCl buffer than in phosphate buffer in the absence of added metals. Both Na+ and Mg2+ facilitate C1, C3, and C4 formation. Na+ also facilitates C3 and C4 formation with the interferon (IFN) response element B. Na+ has little effect on C2 formation with the regulatory region AB, whereas presence of Mg2+ at low concentrations gives a strong C2 signal. A new band, C7, was detected in the presence of 50 or 100 mM Na+. A relatively high Mg2+ concentration is required for C5 formation and Mg2+ interferes with C6 and C7 formation. Cu2+ and Zn2+ markedly inhibit formation of most complexes. However, a new band forms in 0.15 mM Cu2+ when 10 mM Mg2+ was also present. Co2+ enhances C2b and C3 formation and inhibits C4 formation. Mn2+ enhances C3, C1, and C2 formation to different degrees and inhibits C4 formation. At 0.15 and 0.25 mM, Mn2+ generates a new band. In competition experiments between Cu2+ and Mg2+, Cu2+ strongly inhibited Mg2+ effects on complex formation, except that 0.15 or 0.25 mM Cu2+ enhanced the C2a and C2b signals.

Original languageEnglish (US)
Pages (from-to)25-31
Number of pages7
JournalJournal of Interferon Research
Volume11
Issue number1
StatePublished - 1991
Externally publishedYes

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2',5'-Oligoadenylate Synthetase
Nucleic Acid Regulatory Sequences
Carrier Proteins
Buffers
Metals
BALB 3T3 Cells
Tromethamine
Response Elements
Electrophoretic Mobility Shift Assay
Interferons
Genes
Electrophoresis
Phosphates

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

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title = "Effects of metals on the binding of protein factors to the mouse 2',5'- oligoadenylate synthetase ME-12 gene regulatory region",
abstract = "We have previously demonstrated that using mouse 2',5'-oligoadenylate synthetase (2-5A synthetase) gene (ME-12) regulatory region AB as a probe, at least six complexes (C1-C6) can be detected by the mobility shift electrophoresis assay with extracts of murine BALB/c-3T3 cells. The formation of these complexes is greatly influenced by different buffer conditions and the presence of metals. C1 and C4 signals are stronger in Tris-HCl buffer than in phosphate buffer in the absence of added metals. Both Na+ and Mg2+ facilitate C1, C3, and C4 formation. Na+ also facilitates C3 and C4 formation with the interferon (IFN) response element B. Na+ has little effect on C2 formation with the regulatory region AB, whereas presence of Mg2+ at low concentrations gives a strong C2 signal. A new band, C7, was detected in the presence of 50 or 100 mM Na+. A relatively high Mg2+ concentration is required for C5 formation and Mg2+ interferes with C6 and C7 formation. Cu2+ and Zn2+ markedly inhibit formation of most complexes. However, a new band forms in 0.15 mM Cu2+ when 10 mM Mg2+ was also present. Co2+ enhances C2b and C3 formation and inhibits C4 formation. Mn2+ enhances C3, C1, and C2 formation to different degrees and inhibits C4 formation. At 0.15 and 0.25 mM, Mn2+ generates a new band. In competition experiments between Cu2+ and Mg2+, Cu2+ strongly inhibited Mg2+ effects on complex formation, except that 0.15 or 0.25 mM Cu2+ enhanced the C2a and C2b signals.",
author = "Cong Yan and I. Tamm",
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T1 - Effects of metals on the binding of protein factors to the mouse 2',5'- oligoadenylate synthetase ME-12 gene regulatory region

AU - Yan, Cong

AU - Tamm, I.

PY - 1991

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N2 - We have previously demonstrated that using mouse 2',5'-oligoadenylate synthetase (2-5A synthetase) gene (ME-12) regulatory region AB as a probe, at least six complexes (C1-C6) can be detected by the mobility shift electrophoresis assay with extracts of murine BALB/c-3T3 cells. The formation of these complexes is greatly influenced by different buffer conditions and the presence of metals. C1 and C4 signals are stronger in Tris-HCl buffer than in phosphate buffer in the absence of added metals. Both Na+ and Mg2+ facilitate C1, C3, and C4 formation. Na+ also facilitates C3 and C4 formation with the interferon (IFN) response element B. Na+ has little effect on C2 formation with the regulatory region AB, whereas presence of Mg2+ at low concentrations gives a strong C2 signal. A new band, C7, was detected in the presence of 50 or 100 mM Na+. A relatively high Mg2+ concentration is required for C5 formation and Mg2+ interferes with C6 and C7 formation. Cu2+ and Zn2+ markedly inhibit formation of most complexes. However, a new band forms in 0.15 mM Cu2+ when 10 mM Mg2+ was also present. Co2+ enhances C2b and C3 formation and inhibits C4 formation. Mn2+ enhances C3, C1, and C2 formation to different degrees and inhibits C4 formation. At 0.15 and 0.25 mM, Mn2+ generates a new band. In competition experiments between Cu2+ and Mg2+, Cu2+ strongly inhibited Mg2+ effects on complex formation, except that 0.15 or 0.25 mM Cu2+ enhanced the C2a and C2b signals.

AB - We have previously demonstrated that using mouse 2',5'-oligoadenylate synthetase (2-5A synthetase) gene (ME-12) regulatory region AB as a probe, at least six complexes (C1-C6) can be detected by the mobility shift electrophoresis assay with extracts of murine BALB/c-3T3 cells. The formation of these complexes is greatly influenced by different buffer conditions and the presence of metals. C1 and C4 signals are stronger in Tris-HCl buffer than in phosphate buffer in the absence of added metals. Both Na+ and Mg2+ facilitate C1, C3, and C4 formation. Na+ also facilitates C3 and C4 formation with the interferon (IFN) response element B. Na+ has little effect on C2 formation with the regulatory region AB, whereas presence of Mg2+ at low concentrations gives a strong C2 signal. A new band, C7, was detected in the presence of 50 or 100 mM Na+. A relatively high Mg2+ concentration is required for C5 formation and Mg2+ interferes with C6 and C7 formation. Cu2+ and Zn2+ markedly inhibit formation of most complexes. However, a new band forms in 0.15 mM Cu2+ when 10 mM Mg2+ was also present. Co2+ enhances C2b and C3 formation and inhibits C4 formation. Mn2+ enhances C3, C1, and C2 formation to different degrees and inhibits C4 formation. At 0.15 and 0.25 mM, Mn2+ generates a new band. In competition experiments between Cu2+ and Mg2+, Cu2+ strongly inhibited Mg2+ effects on complex formation, except that 0.15 or 0.25 mM Cu2+ enhanced the C2a and C2b signals.

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