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The apical stem-loo...
The apical stem-loop of the hepatitis B virus encapsidation signal folds into a stable tri-loop with two underlying pyrimidine bulges.
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- Flodell, Sara (author)
- Umeå universitet,Institutionen för medicinsk kemi och biofysik
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- Schleucher, Jurgen (author)
- Umeå universitet,Institutionen för medicinsk kemi och biofysik
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- Cromsigt, Jenny (author)
- Umeå universitet,Institutionen för medicinsk kemi och biofysik
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- Ippel, Hans (author)
- Umeå universitet,Institutionen för medicinsk kemi och biofysik
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- Kidd-Ljunggren, Karin (author)
- Lund University,Lunds universitet,Infektionsmedicin,Sektion III,Institutionen för kliniska vetenskaper, Lund,Medicinska fakulteten,Infection Medicine (BMC),Section III,Department of Clinical Sciences, Lund,Faculty of Medicine
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Wijmenga, Sybren (author)
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(creator_code:org_t)
- Oxford University Press (OUP), 2002
- 2002
- English.
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In: Nucleic Acids Research. - : Oxford University Press (OUP). - 0305-1048 .- 1362-4962. ; 30:21, s. 4803-4811
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http://www.ncbi.nlm....
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http://dx.doi.org/10...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Subject headings
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- Reverse transcription of hepatitis B virus (HBV) pregenomic RNA is essential for virus replication. In the first step of this process, HBV reverse transcriptase binds to the highly conserved encapsidation signal, epsilon (epsilon), situated near the 5' end of the pregenome. epsilon has been predicted to form a bulged stem-loop with the apical stem capped by a hexa- loop. After the initial binding to this apical stem- loop, the reverse transcriptase synthesizes a 4 nt primer using the bulge as a template. Here we present mutational and structural data from NMR on the apical stem-loop of epsilon. Application of new isotope-labeling techniques (13C/15N/2H-U-labeling) allowed resolution of many resonance overlaps and an extensive structural data set could be derived. The NMR data show that, instead of the predicted hexa-loop, the apical stem is capped by a stable UGU tri-loop closed by a C-G base pair, followed by a bulged out C. The apical stem contains therefore two unpaired pyrimidines (C1882 and U1889), rather than one as was predicted, spaced by 6 nt. C1882, the 3' neighbour to the G of the loop-closing C-G base pair, is completely bulged out, while U1889 is at least partially intercalated into the stem. Analysis of 205 of our own HBV sequences and 1026 strains from the literature, covering all genotypes, reveals a high degree of conservation of epsilon. In particular, the residues essential for this fold are either totally conserved or show rare non-disruptive mutations. These data strongly indicate that this fold is essential for recognition by the reverse transcriptase.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin -- Infektionsmedicin (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Clinical Medicine -- Infectious Medicine (hsv//eng)
Keyword
- Base Pairing
- Base Sequence
- Genome; Viral
- Genotype
- Hepatitis B virus/classification/enzymology/*genetics
- Humans
- Mutation
- Nuclear Magnetic Resonance; Biomolecular
- Nucleic Acid Conformation
- Pyrimidines/*metabolism
- RNA; Viral/*chemistry/genetics/*metabolism
- RNA-Directed DNA Polymerase/metabolism
- Substrate Specificity
- Uridine/metabolism
- MEDICINE
- MEDICIN
Publication and Content Type
- ref (subject category)
- art (subject category)
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