Dataset Information

Identification of Host factors that Interact with Dengue virus RNA in cell culture

ABSTRACT: Host factors are essential for efficient amplification of Dengue virus (DENV), yet the current knowledge of these cellular proteins remains limited. Here, we used quantitative thiouridine cross-linking mass spectrometry (qTUX-MS) to cross-link proteins to viral RNA during a live DENV infection in cell culture. We identified 79 novel host proteins that have not been previously implicated in DENV infection, and demonstrated the reliability of qTUX-MS by validating a subset of these factors. Analysis of proteome changes revealed critical pathways up- and down-regulated during DENV infection, while the levels of qTUX-MS identified factors remained largely unchanged. Knockdown of HMCES, RBMX and hnRNP F reduced the levels of both intracellular viral RNA and DENV titers, suggesting roles in viral amplification prior to packaging and release. In contrast, knockdown of hnRNP M or NONO caused a dramatic drop in viral titers without impacting viral RNA accumulation, indicating a role downstream of DENV replication. Importantly, none of these five host factors were essential for cell viability or amplification of an unrelated virus, adenovirus, demonstrating that knockdown of these host factors did not reduce cell fitness for viral amplification. Thus, qTUX-MS can be used to identify host factor interactions for any RNA virus.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hepatocyte

DISEASE(S): Dengue Disease

SUBMITTER: Todd Greco

LAB HEAD: Ileana M Cristea

PROVIDER: PXD003593 | Pride | 2016-09-26

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	2013_08_uniprotS_human_DENV.fasta	Fasta
	2014_04_06_TUXMS_DENV_P1-01.raw	Raw
	2014_04_06_TUXMS_DENV_P1-02.raw	Raw
	2014_04_06_TUXMS_DENV_P1-03.raw	Raw
	2014_04_06_qTUXMS_IP_MQresults.zip	Other

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Publications

Identification of RNA Binding Proteins Associated with Dengue Virus RNA in Infected Cells Reveals Temporally Distinct Host Factor Requirements.

Viktorovskaya Olga V OV Greco Todd M TM Cristea Ileana M IM Thompson Sunnie R SR

PLoS neglected tropical diseases 20160824 8

<h4>Background</h4>There are currently no vaccines or antivirals available for dengue virus infection, which can cause dengue hemorrhagic fever and death. A better understanding of the host pathogen interaction is required to develop effective therapies to treat DENV. In particular, very little is known about how cellular RNA binding proteins interact with viral RNAs. RNAs within cells are not naked; rather they are coated with proteins that affect localization, stability, translation and (for v ...[more]

PMID: 27556644

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