Abstract:
:The icosahedral bacteriophage T7 is a 50 MDa double-stranded DNA (dsDNA) virus that infects Escherichia coli. Although there is substantial information on the physical and morphological properties of T7, structural information, based mostly on Raman spectroscopy and cryo-electron microscopy, is limited. Here, we apply the magic-angle spinning (MAS) solid-state NMR (SSNMR) technique to study a uniformly (13)C and (15)N labeled wild-type T7 phage. We describe the details of the large-scale preparation and purification of an isotopically enriched phage sample under fully hydrated conditions, and show a complete (13)C and a near-complete (15)N nucleotide-type specific assignment of the sugar and base moieties in the 40 kbp dsDNA of T7 using two-dimensional (13)C-(13)C and (15)N-(13)C correlation experiments. The chemical shifts are interpreted as reporters of a B-form conformation of the encapsulated dsDNA. While MAS SSNMR was found to be extremely useful in determining the structures of proteins in native-like environments, its application to nucleic acids has lagged behind, leaving a missing (13)C and (15)N chemical shift database. This work therefore expands the (13)C and (15)N database of real B-form DNA systems, and opens routes to characterize more complex nucleic acid systems by SSNMR.
journal_name
J Biomol NMRjournal_title
Journal of biomolecular NMRauthors
Abramov G,Goldbourt Adoi
10.1007/s10858-014-9840-4subject
Has Abstractpub_date
2014-08-01 00:00:00pages
219-30issue
4eissn
0925-2738issn
1573-5001journal_volume
59pub_type
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/BF00211783
更新日期:1995-09-01 00:00:00
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journal_title:Journal of biomolecular NMR
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更新日期:2015-06-01 00:00:00
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journal_title:Journal of biomolecular NMR
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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doi:10.1007/s10858-019-00285-x
更新日期:2019-12-01 00:00:00
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