Ubiquitination of lysine-331 by Kaposi's sarcoma-associated herpesvirus protein K5 targets HFE for lysosomal degradation

Rhodes, David A.; Boyle, Louise H.; Boname, Jessica M.; Lehner, Paul J.; Trowsdale, John

doi:10.1073/pnas.1003421107

Cited by 16 publications

(16 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Published data show that both K3 and K5 are capable of causing increased endocytosis of target proteins, but K5 also targets a sub-set of proteins through altered trafficking from the endoplasmic reticulum or Golgi [11], [48], [49]. Given our finding that K3 or K5 mutated in the tyrosine-based endocytosis motif lost the ability to regulate DC-SIGN and DC-SIGNR, we sought to determine if these lectins were also endocytosed.…”

Section: Resultsmentioning

confidence: 99%

“…Unpublished data, alongside data presented here, indicate however that some synthesized DC-SIGN and DC-SIGNR is able to reach the surface before being targeted for downmodulation by K3 or K5. Interestingly, it has previously been shown that K5 can target the nonclassical MHC I-related molecule HFE both from the cell surface and from the Golgi or a post-Golgi compartment within the cell [49]. K5-associated HFE, in that case, was found to be EndoH insensitive.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Kaposi’s Sarcoma-Associated Herpesvirus K3 and K5 Proteins Down Regulate Both DC-SIGN and DC-SIGNR

et al. 2013

View full text Add to dashboard Cite

Kaposi’s sarcoma-associated herpesvirus (KSHV) is the etiological agent of multicentric Castleman’s disease, primary effusion lymphoma and Kaposi’s sarcoma. In this study, we show that like the C-type lectin DC-SIGN, the closely related DC-SIGNR can also enhance KSHV infection. Following infection, they are both targeted for down modulation and our data indicate that the KSHV MARCH-family ubiquitin ligase K5 is mediating this regulation and subsequent targeting for degradation of DC-SIGN and DC-SIGNR in the context of the virus. The closely related viral K3 protein, is also able to target these lectins in exogenous expressions studies, but only weakly during viral infection. In addition to requiring a functional RING-CH domain, several protein trafficking motifs in the C-terminal region of both K3 and K5 are important in regulation of DC-SIGN and DC-SIGNR. Further exploration of this modulation revealed that DC-SIGN is endocytosed from the cell surface in THP-1 monocytes, but degraded from an internal location with minimal endocytosis in HEK-293 cells. Pull-down data indicate that both K3 and K5 preferentially associate with immature forms of the lectins, mediating their ubiquitylation and degradation. Together, these data emphasize the molecular complexities of K3 and K5, while expanding the repertoire of targets of these two viral proteins.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Kaposi’s Sarcoma-Associated Herpesvirus K3 and K5 Proteins Down Regulate Both DC-SIGN and DC-SIGNR

et al. 2013

View full text Add to dashboard Cite

show abstract

“…More recent evidence suggests a broad role for K5 in various aspects of cellular physiology and homeostasis, including remodeling of endothelial cell junctions through downregulation of VEcadherins, modulation of iron import and export via HFE degradation, inhibition of BMPRII signaling, and increase of monocyte metabolism and proliferation via modulation of the localization-dependent activity of certain receptor tyrosine kinases (22,33,43,54). Interestingly, the latter function can be mediated by K5 mutants lacking an intact RING-CH domain, suggesting E3 ligase activity is not required for certain K5 functions.…”

Section: Discussionmentioning

confidence: 99%

Construction and Manipulation of a New Kaposi's Sarcoma-Associated Herpesvirus Bacterial Artificial Chromosome Clone

Brulois

Chang

Lee

et al. 2012

J Virol

313

455

View full text Add to dashboard Cite

Efficient genetic modification of herpesviruses such as Kaposi's sarcoma-associated herpesvirus (KSHV) has come to rely on bacterial artificial chromosome (BAC) technology. In order to facilitate this approach, we generated a new KSHV BAC clone, called BAC16, derived from the rKSHV.219 virus, which stems from KSHV and Epstein-Barr virus-coinfected JSC1 primary effusion lymphoma (PEL) cells. Restriction enzyme and complete sequencing data demonstrate that the KSHV of JSC1 PEL cells showed a minimal level of sequence variation across the entire viral genome compared to the complete genomic sequence of other KSHV strains. BAC16 not only stably propagated in bothEscherichia coliand mammalian cells without apparent genetic rearrangements, but also was capable of robustly producing infectious virions (∼5 × 107/ml). We also demonstrated the utility of BAC16 by generating deletion mutants of either the K3 or K5 genes, whose products are E3 ligases of the membrane-associated RING-CH (MARCH) family. While previous studies have shown that individual expression of either K3 or K5 results in efficient downregulation of the surface expression of major histocompatibility complex class I (MHC-I) molecules, we found that K5, but not K3, was the primary factor critical for the downregulation of MHC-I surface expression during KSHV lytic reactivation or followingde novoinfection. The data presented here demonstrate the utility of BAC16 for the generation and characterization of KSHV knockout and mutant recombinants and further emphasize the importance of functional analysis of viral genes in the context of the KSHV genome besides the study of individual gene expression.

show abstract

“…Subsequent studies revealed an increasing number of K5 substrates, including the NKT cell ligand CD1d (25); the MHC-Irelated molecule HFE (26); the adhesion molecules ICAM-1 (27,28), PECAM (29), VE-cadherin (30), ALCAM (31), DC-SIGN, and DC-SIGNR (32); the costimulatory molecule B7-2 (27,28); the NK cell-activating ligands, MICA, MICB, and AICL (33); the cellular restriction factor tetherin (34); the cytokine receptor interferon gamma receptor 1 (IFN-␥R1) (35); the plasma membrane t-SNARE syntaxin-4 (36); and the transforming growth factor ␤ (TGF-␤) family member BMPRII (37). In addition, Timms et al recently reported several novel K5 substrates, including 8 verified targets, CD32, CD33, CD99, EPHB4, Plexin A1, PMZL2, Kit (CD117), and IL9R (CD129), and 66 potential new targets, all of which were identified via a quantitative mass spectrometry approach (38).…”

mentioning

confidence: 99%

Kaposi's Sarcoma-Associated Herpesvirus K3 and K5 Ubiquitin E3 Ligases Have Stage-Specific Immune Evasion Roles during Lytic Replication

Brulois

Tóth

Wong

et al. 2014

J Virol

View full text Add to dashboard Cite

The downregulation of immune synapse components such as major histocompatibility complex class I (MHC-I) and ICAM-1 is a common viral immune evasion strategy that protects infected cells from targeted elimination by cytolytic effector functions of the immune system. Kaposi's sarcoma-associated herpesvirus (KSHV) encodes two membrane-bound ubiquitin E3 ligases, called K3 and K5, which share the ability to induce internalization and degradation of MHC-I molecules. Although individual functions of K3 and K5 outside the viral genome are well characterized, their roles during the KSHV life cycle are still unclear. In this study, we individually introduced the amino acid-coding sequences of K3 or K5 into a ⌬K3 ⌬K5 recombinant virus, at either original or interchanged genomic positions. Recombinants harboring coding sequences within the K5 locus showed higher K3 and K5 protein expression levels and more rapid surface receptor downregulation than cognate recombinants in which coding sequences were introduced into the K3 locus. To identify infected cells undergoing K3-mediated downregulation of MHC-I, we employed a novel reporter virus, called red-green-blue-BAC16 (RGB-BAC16), which was engineered to harbor three fluorescent protein expression cassettes: EF1␣-monomeric red fluorescent protein 1 (mRFP1), polyadenylated nuclear RNA promoter (pPAN)-enhanced green fluorescent protein (EGFP), and pK8.1-monomeric blue fluorescent protein (tagBFP), marking latent, immediate early, and late viral gene expression, respectively. Analysis of RGB-derived K3 and K5 deletion mutants showed that while the K5-mediated downregulation of MHC-I was concomitant with pPAN induction, the reduction of MHC-I surface expression by K3 was evident in cells that were enriched for pPAN-driven EGFP high and pK8.1-driven blue fluorescent proteinpositive (BFP ؉ ) populations. These data support the notion that immunoreceptor downregulation occurs by a sequential process wherein K5 is critical during the immediately early phase and K3 plays a significant role during later stages. IMPORTANCEAlthough the roles of K3 and K5 outside the viral genome are well characterized, the function of these proteins in the context of the KSHV life cycle has remained unclear, particularly in the case of K3. This study examined the relative contributions of K3 and K5 to the downregulation of MHC-I during the lytic replication of KSHV. We show that while K5 acts immediately upon entry into the lytic phase, K3-mediated downregulation of MHC-I was evident during later stages of lytic replication. The identification of distinctly timed K3 and K5 activities significantly advances our understanding of KSHV-mediated immune evasion. Crucial to this study was the development of a novel recombinant KSHV, called RGB-BAC16, which facilitated the delineation of stage-specific phenotypes.

show abstract

Ubiquitination of lysine-331 by Kaposi's sarcoma-associated herpesvirus protein K5 targets HFE for lysosomal degradation

Cited by 16 publications

References 35 publications

Kaposi’s Sarcoma-Associated Herpesvirus K3 and K5 Proteins Down Regulate Both DC-SIGN and DC-SIGNR

Kaposi’s Sarcoma-Associated Herpesvirus K3 and K5 Proteins Down Regulate Both DC-SIGN and DC-SIGNR

Construction and Manipulation of a New Kaposi's Sarcoma-Associated Herpesvirus Bacterial Artificial Chromosome Clone

Kaposi's Sarcoma-Associated Herpesvirus K3 and K5 Ubiquitin E3 Ligases Have Stage-Specific Immune Evasion Roles during Lytic Replication

Contact Info

Product

Resources

About