Infectivity of Dengue Virus Serotypes 1 and 2 is Correlated to E Protein Intrinsic Dynamics But Not to Envelope Conformations

Sharma, Kamal; Lim, Xin Ni; Tantirimudalige, Sarala Neomi; Gupta, Anjali; Marzinek, Jan K.; Holdbrook, Daniel A.; Lim, Xin Ying; Bond, Peter J.; Anand, Ganesh S.; Wohland, Thorsten

doi:10.2139/ssrn.3193413

Cited by 1 publication

(2 citation statements)

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“…We used FRET-FCS (44,45) which analyses the fluctuations in FRET efficiency, caused by doubly labeled 5'UAR conformational fluctuations. Unlike FCS, FRET-FCS measurers the proximity ratio (p), which is function related to FRET efficiency (45)(46)(47)(48)(49)(50) and depends on the separation between donor and acceptor but not on the position of the molecules in the observation volume. Thus, the correlation function of the proximity ratio p (Gp) provides information about structural dynamics of the various 5'UAR conformations.…”

Section: Denv2c Protein Facilitates 5'uar/c5'uar 5'uar/3'uar Annealinmentioning

confidence: 99%

“…A value of 1 for β indicates that the system displays normal two-state Arrhenius kinetics, with one discrete energy barrier, while a value of 0 for β indicates a continuum of equal energy barriers. The approach was calibrated as mentioned by Sharma KK et al (49) and effective relaxation time (τ p ) associated with the motion of the 5'UAR hairpin extremities, were determined. Addition of DENV2C to the 5'UAR hairpin resulted in a ~2-fold increase in values of τ p (from 74 ± 21 µs in the absence to 135 ± 28 µs in the presence of DENV2C) ( Fig.…”

Section: Denv2c Protein Facilitates 5'uar/c5'uar 5'uar/3'uar Annealinmentioning

confidence: 99%

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Dengue virus strain 2 capsid protein switches the annealing pathway and reduces the intrinsic dynamics of the conserved 5’ untranslated region

Yong

Raghuvamsi

Anand

et al. 2020

Preprint

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The capsid protein of Dengue Virus strain 2 (DENV2C) is a structural protein with RNA chaperone activity that promotes multiple nucleic acid structural rearrangements, critical for transcription of the single-stranded positive-sense DENV2 genomic RNA. Annealing of the conserved 5' untranslated region (5'UTR) to either its complementary sequence or to the 3' untranslated region (3'UTR) occurs during (+)/(-) ds-RNA formation and (+) RNA circularization, respectively, both essential steps during DENV RNA replication. We investigated the effect of DENV2C on the annealing mechanism of two hairpin structures from the 5'UTR region (21-nt upstream AUG region (5'UAR) and 23-nt capsid-coding hairpin (5'cHP)) to their complementary sequences during (+)/(-) ds-RNA formation and (+) RNA circularization. Using fluorescence spectroscopy, DENV2C was found to switch annealing reactions nucleated mainly through kissing-loop intermediates to stem-stem interactions during (+)/(-) ds-RNA formation while it promotes annealing mainly through kissing-loop interactions during the (+) RNA circularization. Using FRET-FCS and trFRET, we determined that DENV2C exerts RNA chaperone activities by modulating intrinsic dynamics and by reducing the kinetically trapped unfavorable conformations of the 5'UTR sequence. Thus, DENV2C is likely to facilitate genome folding into functional conformations required for replication, playing a role in modulating (+)/(-) ds-RNA formation and (+) RNA circularization. INTRODCTIONDengue fever is transmitted by the bite of an Aedes aegypti or Aedes albopictus mosquito carrying the dengue virus (DENV). DENV has four antigenically-distinct serotypes, DENV1-4, which complicate vaccine development because an effective vaccine should neutralize all four serotypes effectively (1). This is essential because secondary dengue infection tends to cause severe symptoms, such as dengue hemorrhagic fever and can even be fatal (2).DENV2 is a positive-sense single-stranded RNA (ssRNA) virus with an icosahedral structure (T=3). The 50 nm virus has a genome of approximately 10.7 kb, which is translated to a polypeptide that is cleaved into three structural (envelope [E], pre-membrane [prM] and capsid [C]) and seven non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5) (3). The coding region is flanked at both ends by untranslated regions (UTR). The 5′ end has a type I cap structure (m 7 GpppAmp) mediating capdependent translation, but the virus can switch to a noncanonical translation mechanism when

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Section: Denv2c Protein Facilitates 5'uar/c5'uar 5'uar/3'uar Annealinmentioning

confidence: 99%

Section: Denv2c Protein Facilitates 5'uar/c5'uar 5'uar/3'uar Annealinmentioning

confidence: 99%

Dengue virus strain 2 capsid protein switches the annealing pathway and reduces the intrinsic dynamics of the conserved 5’ untranslated region

Yong

Raghuvamsi

Anand

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Infectivity of Dengue Virus Serotypes 1 and 2 is Correlated to E Protein Intrinsic Dynamics But Not to Envelope Conformations

Cited by 1 publication

References 21 publications

Dengue virus strain 2 capsid protein switches the annealing pathway and reduces the intrinsic dynamics of the conserved 5’ untranslated region

Dengue virus strain 2 capsid protein switches the annealing pathway and reduces the intrinsic dynamics of the conserved 5’ untranslated region

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