Tyrosine 27 of the specificity polypeptide of EcoKI can be UV crosslinked to a bromodeoxyuridine-substituted DNA target sequence.
The specificity (S) subunit of the restriction enzyme EcoKI imparts specificity for the sequence AAC(N6)GTGC. Substitution of thymine with bromodeoxyuridine in a 25 bp DNA duplex containing this sequence stimulated UV light-induced covalent crosslinking to the S subunit. Crosslinking occurred only a...
| Main Authors: | , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
1995
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| _version_ | 1797058327305256960 |
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| author | Chen, A Powell, L Dryden, D Murray, N Brown, T |
| author_facet | Chen, A Powell, L Dryden, D Murray, N Brown, T |
| author_sort | Chen, A |
| collection | OXFORD |
| description | The specificity (S) subunit of the restriction enzyme EcoKI imparts specificity for the sequence AAC(N6)GTGC. Substitution of thymine with bromodeoxyuridine in a 25 bp DNA duplex containing this sequence stimulated UV light-induced covalent crosslinking to the S subunit. Crosslinking occurred only at the residue complementary to the first adenine in the AAC sequence, demonstrating a close contact between the major groove at this sequence and the S subunit. Peptide sequencing of a proteolytically-digested, crosslinked complex identified tyrosine 27 in the S subunit as the site of crosslinking. This is consistent with the role of the N-terminal domain of the S subunit in recognizing the AAC sequence. Tyrosine 27 is conserved in the S subunits of the three type I enzymes that share the sequence AA in the trinucleotide component of their target sequence. This suggests that tyrosine 27 may make a similar DNA contact in these other enzymes. |
| first_indexed | 2024-03-06T19:48:55Z |
| format | Journal article |
| id | oxford-uuid:2348b91d-9d4c-4fe0-811c-c7752b8d7b6b |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T19:48:55Z |
| publishDate | 1995 |
| record_format | dspace |
| spelling | oxford-uuid:2348b91d-9d4c-4fe0-811c-c7752b8d7b6b2022-03-26T11:43:32ZTyrosine 27 of the specificity polypeptide of EcoKI can be UV crosslinked to a bromodeoxyuridine-substituted DNA target sequence.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:2348b91d-9d4c-4fe0-811c-c7752b8d7b6bEnglishSymplectic Elements at Oxford1995Chen, APowell, LDryden, DMurray, NBrown, TThe specificity (S) subunit of the restriction enzyme EcoKI imparts specificity for the sequence AAC(N6)GTGC. Substitution of thymine with bromodeoxyuridine in a 25 bp DNA duplex containing this sequence stimulated UV light-induced covalent crosslinking to the S subunit. Crosslinking occurred only at the residue complementary to the first adenine in the AAC sequence, demonstrating a close contact between the major groove at this sequence and the S subunit. Peptide sequencing of a proteolytically-digested, crosslinked complex identified tyrosine 27 in the S subunit as the site of crosslinking. This is consistent with the role of the N-terminal domain of the S subunit in recognizing the AAC sequence. Tyrosine 27 is conserved in the S subunits of the three type I enzymes that share the sequence AA in the trinucleotide component of their target sequence. This suggests that tyrosine 27 may make a similar DNA contact in these other enzymes. |
| spellingShingle | Chen, A Powell, L Dryden, D Murray, N Brown, T Tyrosine 27 of the specificity polypeptide of EcoKI can be UV crosslinked to a bromodeoxyuridine-substituted DNA target sequence. |
| title | Tyrosine 27 of the specificity polypeptide of EcoKI can be UV crosslinked to a bromodeoxyuridine-substituted DNA target sequence. |
| title_full | Tyrosine 27 of the specificity polypeptide of EcoKI can be UV crosslinked to a bromodeoxyuridine-substituted DNA target sequence. |
| title_fullStr | Tyrosine 27 of the specificity polypeptide of EcoKI can be UV crosslinked to a bromodeoxyuridine-substituted DNA target sequence. |
| title_full_unstemmed | Tyrosine 27 of the specificity polypeptide of EcoKI can be UV crosslinked to a bromodeoxyuridine-substituted DNA target sequence. |
| title_short | Tyrosine 27 of the specificity polypeptide of EcoKI can be UV crosslinked to a bromodeoxyuridine-substituted DNA target sequence. |
| title_sort | tyrosine 27 of the specificity polypeptide of ecoki can be uv crosslinked to a bromodeoxyuridine substituted dna target sequence |
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