Host factors interacting with the dengue virus (DENV) 39 UTR are involved in virus replication, but their roles remain poorly understood. We used RNA affinity capture and mass spectrometry to identify p100 as a host cellular protein associated with the DENV 39 UTR. By using RNA immunoprecipitation and confocal immunofluorescence analysis we demonstrated an interaction between p100 and the 39 UTR in DENV-infected cells. We identified the A4 region (the extensive stem-loop structure at the 39 end) as the binding site of p100 by studying deletion mutants. p100 knockdown specifically reduced the levels of viral RNA and viral protein in DENV-infected cells. Furthermore, downregulation of p100 reduced the expression of a heterologously expressed luciferase-39 UTR(DENV) mRNA in an A4-dependent manner, confirming the binding data and the effects of p100 knockdown on viral replication. These results provide evidence that p100 interacts with the 39 UTR of DENV and is required for normal DENV replication.
This paper discusses the kinetic fractionation, composition and distribution characteristics of carbon and hydrogen isotopes for various alkane gases formed in different environments, by different mechanisms and from different sources in nature.It is demonstrated that the biodegradation or thermodegradation of complex high-molecule sedimentary organic material can form microbial gas or thermogenic gas. The δ 13 C 1 value ranges from −110‰ to −50‰ for microbial gases but from −50‰ to −35‰ (even heavier) for thermogenic gases. Controlled by the kinetic isotope fractionation, both microbial and thermogenic gases have δ 13 C and δ D values characterized by normal distribution, i.e. δ 13 C 1 < δ 13 C 2 < δ 13 C 3 < δ 13 C 4 and δ D CH 4 < δ D C 2 H 6 < δ D C 3 H 8 <δ D C 4 H 10 , and by a positive correlation between the δ 13 C and δ D values. Simple carbon-bearing molecules (CH 4 , CO and CO 2 ) can form abiogenic alkane gases via polymerization in the abiological chemical process in nature, with δ 13 C 1 heavier than −30‰.Moreover, controlled by the kinetic isotope fractionation, abiogenic alkane gases are characterized by a reverse distribution of δ 13 C values and a normal trend of δ D values, namely δ 13 C 1 > δ 13 C 2 > δ 13 C 3 > δ 13 C 4 and δ D CH 4 <δ D C 2 H 6 < δ D C 3 H 8 < δ D C 4 H 10 . The δ 13 C values and δ D values are negatively correlated. Natural gases from 26 commercial gas wells distributed in the Xujiaweizi and Yingshan-Miaotaizi faulted depressions in the Songliao Basin, China, show δ 13 C 1 values ranging from −30.5‰ to −16.7‰ with a very narrow δ D range between −203‰-−196‰. These gases are characterized by a reverse distribution of δ 13 C values but a normal distribution of δ D values, and a negative correlation between their δ 13 C and δ D values, indicating an abiological origin. The present study has revealed that abiogenic hydrocarbons not only exist in nature but also can make significant contribution to commercial gas reserviors. It is estimated that the reserve volume of alkane gases with abiogenic characteristics in these 26 gas wells in the Songliao Basin is over 500×10 8 m 3 . The prospecting practice in the Songliao Basin has demonstrated that abiogenic alkane gases are of a promising resource, and it provides an example for the investigation of and search for abiogenic commercial natural gases worldwide. generation mechanism of natural gases, kinetic isotope fractionation, the Songliao Basin of China, commercial abiogenic hydrocarbon gaspool
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