Interaction of psoralen-derivatized oligodeoxyribonucleoside methylphosphonates with single-stranded DNA

Bok L. Lee, Akira Murakami, Kathleen Blake, Shwu Bin Lin, Paul S. Miller

Research output: Contribution to journalArticle

Abstract

Oligodeoxyribonucleoside methylphosphonates derivatized at the 5′ end with 4′-(amino-alkyl)-4,5′,8-trimethylpsoralen were prepared. The interaction of these psoralen-derivatized methylphosphonate oligomers with synthetic single-stranded DNAs 35 nucleotides in length was studied. Irradiation of a solution containing the 35-mer and its complementary methylphosphonate oligomer at 365 nm gave a cross-linked duplex produced by cycloaddition between the psoralen pyrone ring of the derivatized me′ thylphosphonate oligomer and a thymine base of the DNA. Photoadduct formation could be reversed by irradiation at 254 nm. The rate and extent of cross-linking were dependent upon the length of the aminoalkyl linker between the trimethylpsoralen group and the 5′ end of the methylphosphonate oligomer. Methylphosphonate oligomers derivatized with 4′-[[N-(2-aminoethyl)amino] methyl]-4,5′,8-trimethylpsoralen gave between 70% and 85% cross-linked product when irradiated for 20 min at 4°C. Further irradiation did not increase cross-linking, and preirradiation of the psoralen-derivatized methylphosphonate oligomer at 365 nm reduced or prevented cross-linking. These results suggest that the methylphosphonate oligomers undergo both cross-linking and deactivation reactions when irradiated at 365 nm. The extent of cross-linking increased up to 10 μM oligomer concentration and dramatically decreased at temperatures above the estimated Tm of the methylphosphonate oligomer-DNA duplex. The cross-linking reaction was dependent upon the fidelity of base-pairing interactions between the methylphosphonate oligomers and the single-stranded DNA. Noncomplementary oligomers did not cross-link, and the extent of cross-linking of oligomers containing varying numbers of noncomplementary bases was greatly diminished or eliminated. The extent and sequence specificity of photoinduced cross-linking, combined with the known ability of methylphosphonate oligomers to be taken up by living cells, suggest that psoralen-derivatized oligonucleoside methylphosphonates may be useful probes of cellular gene expression.

Original languageEnglish (US)
Pages (from-to)3197-3203
Number of pages7
JournalBiochemistry®
Volume27
Issue number9
StatePublished - 1988

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Ficusin
Single-Stranded DNA
Oligomers
Trioxsalen
Irradiation
dideoxyribonucleoside methylphosphonates
methylphosphonic acid
Pyrones
Thymine
DNA
Cross Reactions
Cycloaddition Reaction
Base Pairing
Cycloaddition
Nucleotides
Gene expression

ASJC Scopus subject areas

  • Biochemistry

Cite this

Lee, B. L., Murakami, A., Blake, K., Lin, S. B., & Miller, P. S. (1988). Interaction of psoralen-derivatized oligodeoxyribonucleoside methylphosphonates with single-stranded DNA. Biochemistry®, 27(9), 3197-3203.

Interaction of psoralen-derivatized oligodeoxyribonucleoside methylphosphonates with single-stranded DNA. / Lee, Bok L.; Murakami, Akira; Blake, Kathleen; Lin, Shwu Bin; Miller, Paul S.

In: Biochemistry®, Vol. 27, No. 9, 1988, p. 3197-3203.

Research output: Contribution to journalArticle

Lee, BL, Murakami, A, Blake, K, Lin, SB & Miller, PS 1988, 'Interaction of psoralen-derivatized oligodeoxyribonucleoside methylphosphonates with single-stranded DNA', Biochemistry®, vol. 27, no. 9, pp. 3197-3203.
Lee, Bok L. ; Murakami, Akira ; Blake, Kathleen ; Lin, Shwu Bin ; Miller, Paul S. / Interaction of psoralen-derivatized oligodeoxyribonucleoside methylphosphonates with single-stranded DNA. In: Biochemistry®. 1988 ; Vol. 27, No. 9. pp. 3197-3203.
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abstract = "Oligodeoxyribonucleoside methylphosphonates derivatized at the 5′ end with 4′-(amino-alkyl)-4,5′,8-trimethylpsoralen were prepared. The interaction of these psoralen-derivatized methylphosphonate oligomers with synthetic single-stranded DNAs 35 nucleotides in length was studied. Irradiation of a solution containing the 35-mer and its complementary methylphosphonate oligomer at 365 nm gave a cross-linked duplex produced by cycloaddition between the psoralen pyrone ring of the derivatized me′ thylphosphonate oligomer and a thymine base of the DNA. Photoadduct formation could be reversed by irradiation at 254 nm. The rate and extent of cross-linking were dependent upon the length of the aminoalkyl linker between the trimethylpsoralen group and the 5′ end of the methylphosphonate oligomer. Methylphosphonate oligomers derivatized with 4′-[[N-(2-aminoethyl)amino] methyl]-4,5′,8-trimethylpsoralen gave between 70{\%} and 85{\%} cross-linked product when irradiated for 20 min at 4°C. Further irradiation did not increase cross-linking, and preirradiation of the psoralen-derivatized methylphosphonate oligomer at 365 nm reduced or prevented cross-linking. These results suggest that the methylphosphonate oligomers undergo both cross-linking and deactivation reactions when irradiated at 365 nm. The extent of cross-linking increased up to 10 μM oligomer concentration and dramatically decreased at temperatures above the estimated Tm of the methylphosphonate oligomer-DNA duplex. The cross-linking reaction was dependent upon the fidelity of base-pairing interactions between the methylphosphonate oligomers and the single-stranded DNA. Noncomplementary oligomers did not cross-link, and the extent of cross-linking of oligomers containing varying numbers of noncomplementary bases was greatly diminished or eliminated. The extent and sequence specificity of photoinduced cross-linking, combined with the known ability of methylphosphonate oligomers to be taken up by living cells, suggest that psoralen-derivatized oligonucleoside methylphosphonates may be useful probes of cellular gene expression.",
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N2 - Oligodeoxyribonucleoside methylphosphonates derivatized at the 5′ end with 4′-(amino-alkyl)-4,5′,8-trimethylpsoralen were prepared. The interaction of these psoralen-derivatized methylphosphonate oligomers with synthetic single-stranded DNAs 35 nucleotides in length was studied. Irradiation of a solution containing the 35-mer and its complementary methylphosphonate oligomer at 365 nm gave a cross-linked duplex produced by cycloaddition between the psoralen pyrone ring of the derivatized me′ thylphosphonate oligomer and a thymine base of the DNA. Photoadduct formation could be reversed by irradiation at 254 nm. The rate and extent of cross-linking were dependent upon the length of the aminoalkyl linker between the trimethylpsoralen group and the 5′ end of the methylphosphonate oligomer. Methylphosphonate oligomers derivatized with 4′-[[N-(2-aminoethyl)amino] methyl]-4,5′,8-trimethylpsoralen gave between 70% and 85% cross-linked product when irradiated for 20 min at 4°C. Further irradiation did not increase cross-linking, and preirradiation of the psoralen-derivatized methylphosphonate oligomer at 365 nm reduced or prevented cross-linking. These results suggest that the methylphosphonate oligomers undergo both cross-linking and deactivation reactions when irradiated at 365 nm. The extent of cross-linking increased up to 10 μM oligomer concentration and dramatically decreased at temperatures above the estimated Tm of the methylphosphonate oligomer-DNA duplex. The cross-linking reaction was dependent upon the fidelity of base-pairing interactions between the methylphosphonate oligomers and the single-stranded DNA. Noncomplementary oligomers did not cross-link, and the extent of cross-linking of oligomers containing varying numbers of noncomplementary bases was greatly diminished or eliminated. The extent and sequence specificity of photoinduced cross-linking, combined with the known ability of methylphosphonate oligomers to be taken up by living cells, suggest that psoralen-derivatized oligonucleoside methylphosphonates may be useful probes of cellular gene expression.

AB - Oligodeoxyribonucleoside methylphosphonates derivatized at the 5′ end with 4′-(amino-alkyl)-4,5′,8-trimethylpsoralen were prepared. The interaction of these psoralen-derivatized methylphosphonate oligomers with synthetic single-stranded DNAs 35 nucleotides in length was studied. Irradiation of a solution containing the 35-mer and its complementary methylphosphonate oligomer at 365 nm gave a cross-linked duplex produced by cycloaddition between the psoralen pyrone ring of the derivatized me′ thylphosphonate oligomer and a thymine base of the DNA. Photoadduct formation could be reversed by irradiation at 254 nm. The rate and extent of cross-linking were dependent upon the length of the aminoalkyl linker between the trimethylpsoralen group and the 5′ end of the methylphosphonate oligomer. Methylphosphonate oligomers derivatized with 4′-[[N-(2-aminoethyl)amino] methyl]-4,5′,8-trimethylpsoralen gave between 70% and 85% cross-linked product when irradiated for 20 min at 4°C. Further irradiation did not increase cross-linking, and preirradiation of the psoralen-derivatized methylphosphonate oligomer at 365 nm reduced or prevented cross-linking. These results suggest that the methylphosphonate oligomers undergo both cross-linking and deactivation reactions when irradiated at 365 nm. The extent of cross-linking increased up to 10 μM oligomer concentration and dramatically decreased at temperatures above the estimated Tm of the methylphosphonate oligomer-DNA duplex. The cross-linking reaction was dependent upon the fidelity of base-pairing interactions between the methylphosphonate oligomers and the single-stranded DNA. Noncomplementary oligomers did not cross-link, and the extent of cross-linking of oligomers containing varying numbers of noncomplementary bases was greatly diminished or eliminated. The extent and sequence specificity of photoinduced cross-linking, combined with the known ability of methylphosphonate oligomers to be taken up by living cells, suggest that psoralen-derivatized oligonucleoside methylphosphonates may be useful probes of cellular gene expression.

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