Localization and sub-cellular shuttling of HTLV-1 tax with the miRNA machinery.
The innate ability of the human cell to silence endogenous retroviruses through RNA sequences encoding microRNAs, suggests that the cellular RNAi machinery is a major means by which the host mounts a defense response against present day retroviruses. Indeed, cellular miRNAs target and hybridize to specific sequences of both HTLV-1 and HIV-1 viral transcripts. However, much like the variety of host immune responses to retroviral infection, the virus itself contains mechanisms that assist in the evasion of viral inhibition through control of the cellular RNAi pathway. Retroviruses can hijack both the enzymatic and catalytic components of the RNAi pathway, in some cases to produce novel viral miRNAs that can either assist in active viral infection or promote a latent state. Here, we show that HTLV-1 Tax contributes to the dysregulation of the RNAi pathway by altering the expression of key components of this pathway. A survey of uninfected and HTLV-1 infected cells revealed that Drosha protein is present at lower levels in all HTLV-1 infected cell lines and in infected primary cells, while other components such as DGCR8 were not dramatically altered. We show colocalization of Tax and Drosha in the nucleus in vitro as well as coimmunoprecipitation in the presence of proteasome inhibitors, indicating that Tax interacts with Drosha and may target it to specific areas of the cell, namely, the proteasome. In the presence of Tax we observed a prevention of primary miRNA cleavage by Drosha. Finally, the changes in cellular miRNA expression in HTLV-1 infected cells can be mimicked by the add back of Drosha or the addition of antagomiRs against the cellular miRNAs which are downregulated by the virus.
Department of Molecular and Microbiology, National Center for Biodefense & Infectious Diseases, George Mason University, Manassas, Virginia, United States of America.
Rachel
R
Van Duyne
Irene
I
Guendel
Zachary
Z
Klase
Aarthi
A
Narayanan
William
W
Coley
Elizabeth
E
Jaworski
Jessica
J
Roman
Anastas
A
Popratiloff
Renaud
R
Mahieux
Kylene
K
Kehn-Hall
Fatah
F
Kashanchi
D002467:Cell Nucleus / Q000378:metabolism; D015536:Down-Regulation; D016356:Gene Products, tax / Q000378:metabolism*; D015490:HTLV-I Infections / Q000821:virology; D015368:Human T-lymphotropic virus 1 / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D011506:Proteins / Q000378:metabolism; D034622:RNA Interference; D016601:RNA-Binding Proteins; D043244:Ribonuclease III / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D014779:Virus Replication
C474043:DGCR8 protein, human; D016356:Gene Products, tax; D035683:MicroRNAs; D011506:Proteins; D016601:RNA-Binding Proteins; C517918:DROSHA protein, human; D043244:Ribonuclease III