Bio

Honors & Awards


  • Elizabeth Nash Memorial Fellowship, Cystic Fibrosis Research Inc. (CFRI) (2013-2014)
  • Peer Mentor Award, University of Wisconsin-Madison (2010)
  • Vilas Travel Grant, University of Wisconsin-Madison (2009 and 2010)

Professional Education


  • Doctor of Philosophy, University of Wisconsin Madison (2011)
  • MS, Yonsei University, Biochemistry (2003)
  • BS, Yonsei University, Biochemistry (2001)

Stanford Advisors


Publications

Journal Articles


  • Ubiquitin-Independent Proteasomal Degradation of Tumor Suppressors by Human Cytomegalovirus pp71 Requires the 19S Regulatory Particle JOURNAL OF VIROLOGY Winkler, L. L., Hwang, J., Kalejta, R. F. 2013; 87 (8): 4665-4671

    Abstract

    Proteasomes generally degrade substrates tagged with polyubiquitin chains. In rare cases, however, proteasomes can degrade proteins without prior ubiquitination. For example, the human cytomegalovirus (HCMV) pp71 protein induces the proteasome-dependent, ubiquitin-independent degradation of the retinoblastoma (Rb) and Daxx proteins. These transcriptional corepressors and tumor suppressors inhibit the expression of cellular or viral genes that are required for efficient viral replication. Proteasomes are composed of a 20S catalytic core with or without one or two activator complexes, of which there are four different types. Here, we show that only one of these activators, the 19S regulatory particle that normally participates in ubiquitin-dependent protein degradation, is required for pp71-mediated degradation of Rb and Daxx. We report the unique use of a well-established route of substrate delivery to the proteasome by a viral protein to promote infection.

    View details for DOI 10.1128/JVI.03301-12

    View details for Web of Science ID 000316671000047

    View details for PubMedID 23408605

  • Ubiquitin-independent proteasomal degradation during oncogenic viral infections BIOCHIMICA ET BIOPHYSICA ACTA-REVIEWS ON CANCER Hwang, J., Winkler, L., Kalejta, R. F. 2011; 1816 (2): 147-157

    Abstract

    Most eukaryotic proteins destined for imminent destruction are first tagged with a chain of ubiquitin molecules and are subsequently dismantled by the proteasome. Ubiquitin-independent degradation of substrates by the proteasome, however, also occurs. The number of documented proteasome-dependent, ubiquitin-independent degradation events remains relatively small but continues to grow. Proteins involved in oncogenesis and tumor suppression make up the majority of the known cases for this type of protein destruction. Provocatively, viruses with confirmed or suspected oncogenic properties are also prominent participants in the pantheon of ubiquitin-independent proteasomal degradation events. In this review, we identify and describe examples of proteasome-dependent, ubiquitin-independent protein degradation that occur during tumor virus infections, speculate why this type of protein destruction may be preferred during oncogenesis, and argue that this uncommon type of protein turnover represents a prime target for antiviral and anticancer therapeutics.

    View details for DOI 10.1016/j.bbcan.2011.05.005

    View details for Web of Science ID 000297882300005

    View details for PubMedID 21664948

  • In vivo analysis of protein sumoylation induced by a viral protein: Detection of HCMV pp71-induced Daxx sumoylation METHODS Hwang, J., Kalejta, R. F. 2011; 55 (2): 160-165

    Abstract

    Small ubiquitin-like modifiers (SUMOs) are covalently conjugated to target proteins to regulate numerous biological processes, including subcellular localization, protein-protein interactions, and transactivational activities. While the majority of identified SUMO targets are cellular proteins, SUMO modified viral proteins have also been identified. In addition, there are a growing number of examples where viruses alter the sumoylation status of host cell proteins. Work from our laboratory has previously demonstrated that the human cytomegalovirus (HCMV) virion tegument protein pp71 binds to Daxx, a cellular transcriptional co-repressor, and promotes its sumoylation. Here we describe the in vivo techniques used to detect pp71-induced sumoylation of Daxx in a cotransfection system as well as the endogenous SUMO modified form of Daxx in HCMV-infected cells. The approaches we describe can be easily adapted to infections with other viruses and for the detection of sumoylation of other proteins.

    View details for DOI 10.1016/j.ymeth.2011.07.004

    View details for Web of Science ID 000296933800009

    View details for PubMedID 21816224

  • Elongin B-Mediated Epigenetic Alteration of Viral Chromatin Correlates with Efficient Human Cytomegalovirus Gene Expression and Replication MBIO Hwang, J., Saffert, R. T., Kalejta, R. F. 2011; 2 (2)

    Abstract

    Elongins B and C are members of complexes that increase the efficiency of transcriptional elongation by RNA polymerase II (RNAPII) and enhance the monoubiquitination of histone H2B, an epigenetic mark of actively transcribed genes. Here we show that, in addition to its role in facilitating transcription of the cellular genome, elongin B also enhances gene expression from the double-stranded DNA genome of human cytomegalovirus (HCMV), a pathogenic herpesvirus. Reducing the level of elongin B by small interfering RNA- or short hairpin RNA-mediated knockdown decreased viral mRNA expression, viral protein accumulation, viral DNA replication, and infectious virion production. Chromatin immunoprecipitation analysis indicated viral genome occupancy of the elongating form of RNAPII, and monoubiquitinated histone H2B was reduced in elongin B-deficient cells. These data suggest that, in addition to the previously documented epigenetic regulation of transcriptional initiation, HCMV also subverts cellular elongin B-mediated epigenetic mechanisms for enhancing transcriptional elongation to enhance viral gene expression and virus replication.The genetic and epigenetic control of transcription initiation at both cellular and viral promoters is well documented. Recently, the epigenetic modification of histone H2B monoubiquitination throughout the bodies of cellular genes has been shown to enhance the elongation of RNA polymerase II-initiated transcripts. Mechanisms that might control the elongation of viral transcripts are less well studied. Here we show that, as with cellular genes, elongin B-mediated monoubiquitination of histone H2B also facilitates the transcriptional elongation of human cytomegalovirus genes. This and perhaps other epigenetic markings of actively transcribed regions may help in identifying viral genes expressed during in vitro latency or during natural infections of humans. Furthermore, this work identifies a novel, tractable model system to further study the regulation of transcriptional elongation in living cells.

    View details for DOI 10.1128/mBio.00023-11

    View details for Web of Science ID 000296843700003

    View details for PubMedID 21447700

  • Human Cytomegalovirus Protein pp71 Induces Daxx SUMOylation JOURNAL OF VIROLOGY Hwang, J., Kalejta, R. F. 2009; 83 (13): 6591-6598

    Abstract

    Proteins that participate in a diverse array of cellular processes can be modified covalently and reversibly on lysine residues by the small ubiquitin-like modifier proteins termed SUMOs. In some instances, such modification profoundly affects protein function, but the biological significance of many SUMOylation events remains unknown. Protein SUMOylation is modulated during many viral infections. Here we demonstrate that the human cytomegalovirus (HCMV) pp71 protein promotes the SUMOylation of its cellular substrate, Daxx. A component of promyelocytic leukemia nuclear bodies, Daxx is a transcriptional corepressor that silences the expression of viral immediate-early (IE) genes at the start of both lytic and quiescent HCMV infections. pp71 is a tegument component delivered directly to cells by infecting HCMV virions. At the start of lytic infections, it travels to the nucleus and stimulates viral IE gene expression by displacing the chromatin remodeling protein ATRX from Daxx and by mediating Daxx degradation through a rare ubiquitin-independent, proteasome-dependent process. Here we report that pp71 also substantially increases the basal level of SUMOylated Daxx observed in cells. To date, consequences of Daxx SUMOylation have not been observed for cellular promoters, and we detected no qualitative change in viral IE gene expression in the absence of pp71-induced Daxx SUMOylation. Thus, while pp71 enhances the basal level of SUMOylated Daxx, the role that this modification plays in regulating Daxx activity in uninfected or HCMV-infected cells remains an enigma.

    View details for DOI 10.1128/JVI.02639-08

    View details for Web of Science ID 000267354100026

    View details for PubMedID 19369322

  • Proteasome-dependent, ubiquitin-independent degradation of Daxx by the viral pp71 protein in human cytomegalovirus-infected cells VIROLOGY Hwang, J., Kalejta, R. F. 2007; 367 (2): 334-338

    Abstract

    The cellular Daxx protein represses human cytomegalovirus (HCMV) gene expression from the major immediate early promoter. HCMV prevents Daxx-mediated silencing during lytic infection by delivering the viral pp71 tegument protein to the nucleus, where pp71 binds to and induces the proteasomal degradation of Daxx. In this study, we show that a functional ubiquitin pathway is not required for the proteasomal degradation of the endogenous Daxx protein by tegument-delivered pp71 in HCMV-infected cells, demonstrating that the pp71-mediated degradation of Daxx occurs through a proteasome-dependent, ubiquitin-independent pathway.

    View details for DOI 10.1016/j.virol.2007.05.037

    View details for Web of Science ID 000250162900010

    View details for PubMedID 17590404

  • TIP120A associates with cullins and modulates ubiquitin ligase activity JOURNAL OF BIOLOGICAL CHEMISTRY Min, K. W., Hwang, J. W., Lee, J. S., Park, Y., Tamura, T., Yoon, J. B. 2003; 278 (18): 15905-15910

    Abstract

    The cullin-containing ubiquitin-protein isopeptide ligases (E3s) play an important role in regulating the abundance of key proteins involved in cellular processes such as cell cycle and cytokine signaling. They have multisubunit modular structures in which substrate recognition and the catalysis of ubiquitination are carried out by distinct polypeptides. In a search for proteins involved in regulation of cullin-containing E3 ubiquitin ligases we immunopurified CUL4B-containing complex from HeLa cells and identified TIP120A as an associated protein by mass spectrometry. Immunoprecipitation of cullins revealed that all cullins tested specifically interacted with TIP120A. Reciprocal immunoaffinity purification of TIP120A confirmed the stable interaction of TIP120A with cullin family proteins. TIP120A formed a complex with CUL1 and Rbx1, but interfered with the binding of Skp1 and F-box proteins to CUL1. TIP120A greatly reduced the ubiquitination of phosphorylated IkappaBalpha by SCF(beta-TrCP) ubiquitin ligase. These results suggest that TIP120A functions as a negative regulator of SCF E3 ubiquitin ligases and may modulate other cullin ligases in a similar fashion.

    View details for DOI 10.1074/jbc.M213070200

    View details for Web of Science ID 000182680000057

    View details for PubMedID 12609982

  • TIP120A associates with unneddylated cullin 1 and regulates its neddylation FEBS LETTERS Hwang, J. W., Min, K. W., Tamura, T., Yoon, J. B. 2003; 541 (1-3): 102-108

    Abstract

    The cullin-containing E3 ubiquitin ligases play an important role in regulating the abundance of key proteins involved in cellular processes such as cell cycle and cytokine signaling. We recently identified TIP120A as a cullin-interacting protein and found that TIP120A functions as a negative regulator of a ubiquitin ligase by interfering with the binding of Skp1 and an F box protein to CUL1. Here we show that TIP120A binds to the unneddylated CUL1 but not the neddylated one. The association of TIP120A with CUL1 requires both the N-terminal stalk and the C-terminal globular domain of CUL1. TIP120A efficiently inhibits neddylation of CUL1 but does not affect substrate-independent ubiquitination by CUL1/Rbx1, implying that it blocks the access of Nedd8 to the conjugation site but does not interfere with the interaction of the ubiquitin-conjugating enzyme with Rbx1. Our data suggest that the association/dissociation of TIP120A coupled to neddylation/deneddylation of CUL1 may play an important role in assembly and disassembly of Skp1-Cdc53/cullin-F box ubiquitin ligases.

    View details for DOI 10.1016/S0014-5793(03)00321-1

    View details for Web of Science ID 000182481600020

    View details for PubMedID 12706828

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