Janet Douglas, Ph.D.

Viruses and their hosts are in a constant evolutionary arms race to survive.  Studying this intricate battle provides us with new insights into understanding anti-viral immunity and new targets for developing anti-viral therapies. My research focuses on the interplay between the host’s defense mechanism to prevent viral release from cells and the various viral countermeasures that have co-evolved. 

A major aspect of my work involves characterizing the mechanism of how the HIV-1 protein Vpu counteracts the host restriction factor BST-2/Tetherin to allow efficient viral egress. We are specifically investigating how Vpu and BST-2 interact and how Vpu uses the host’s own ubiquitin system (for normal protein trafficking and degradation) to circumvent BST-2. In addition, the lab is employing various screening methodologies to identify additional host factors that may be necessary for BST-2 function and/or adaptors important for Vpu’s ability to downmodulate BST-2. 

The viral release restriction imposed by BST-2 is not limited to HIV, but may represent an innate immune mechanism that functions to limit the spread of many, if not all enveloped viruses. The filovirus family is composed of both Marburg and Ebola viruses, which are negative-strand RNA viruses that encode only seven ORFs. Despite their simplicity, human infection results in highly transmissible hemorrhagic fevers with mortality rates approaching 90%. Currently there are no treatments or vaccines for Ebola virus. Like HIV, Ebola virus appears to have evolved a countermeasure to BST-2 in the form of its envelope protein GP. Therefore, we are extending our studies to determine how Ebola virus GP is able to overcome the BST-2 restriction. The GP/BST-2 interaction has the potential to be a new target for the design of effective anti-Ebola strategies.

Although it is becoming increasingly apparent that BST-2 functions as a broad spectrum, antiviral innate immune mechanism, there are some enveloped viruses, such as the flaviviruses that bud from cells quite differently than HIV and Ebola and therefore may avoid the BST-2 restriction. My lab is also interested in determining whether a member of the flaviviruses, Dengue is also restricted by BST-2 and if so, if it has evolved a counteracting mechanism.

Biography

After receiving a B.S. in biology at Eckerd College, Janet entered graduate school at the University of Tennessee in Memphis where she received her Ph.D. in 1995 for characterizing the immortalizing and transforming potential of the Adenovirus E1A gene. As a postdoctoral fellow in Dr. Victor Garcia’s laboratory at St. Jude Children’s Hospital in Memphis, TN she developed an HIV-based lentiviral gene delivery system for the transduction of non-dividing human lymphocytes and hematopoietic stem cells. This foray into gene therapy led her into more applied biology, which she pursued at the biotechnology company Systemix in Palo Alto, CA. Here, she extended her work on gene therapy to include a humanized mouse model for assessing the efficacy of lentiviral vectors. Keeping her focus on virology she left the field of gene therapy in 2001 and began working in drug discovery for Gilead Sciences in Foster City, CA.  There, she participated in a multi-disciplinary team of scientists attempting to identify novel small molecule inhibitors of respiratory syncytial virus (RSV) fusion and later inhibitors of HIV-1 reverse transcriptase. In 2004 she returned to academic science and began working with Dr. Ashlee Moses at the Vaccine and Gene Therapy Institute (VGTI) at OHSU where they investigated the cellular transformation mechanisms induced by KSHV/HHV-8 infection, as well as determining how the HIV-1 accessory protein Vpu functions to enhance the release of viral particles by counteracting the host immunomodulatory protein BST-2. Currently, as an assistant scientist at the VGTI she is continuing her research on the host’s viral egress restriction and has extended it to other viruses.

Publications

1: Gustin JK, Douglas JL, Bai Y, Moses AV. Ubiquitination of BST-2 protein by HIV-1 Vpu protein does not require lysine, serine, or threonine residues within the BST-2 cytoplasmic domain. J Biol Chem. 2012 Apr 27; 287(18): 14837-50. PMCID  PMC3340234

2: Gustin JK, Moses AV, Früh K, Douglas JL. Viral takeover of the host ubiquitin system. Front Microbiol. 2011; 2: 161. PMCID: PMC3147166

3: Douglas J. L., J. K. Gustin, A. V. Moses, B. J. Dezube, and L Pantanowitz.  2010. Kaposi Sarcoma pathogenesis:  a triad of viral infection, oncogenesis, and chronic inflammation. Trans Biomed. 2010 1(2): 1-15. PMCID: PMC3472629

4: Douglas JL, Gustin JK, Viswanathan K, Mansouri M, Moses AV, Früh K. The great escape: viral strategies to counter BST-2/tetherin. PLoS Pathog. 2010 May 13; 6(5): e1000913. Review. PMCID: PMC2869331

5: Mansouri M, Viswanathan K, Douglas JL, Hines J, Gustin J, Moses AV, Früh K. Molecular mechanism of BST2/tetherin downregulation by K5/MIR2 of Kaposi's sarcoma-associated herpesvirus. J Virol. 2009 Oct; 83(19): 9672-81. PMCID: PMC2748026

6: Douglas JL, Viswanathan K, McCarroll MN, Gustin JK, Früh K, Moses AV. Vpu directs the degradation of the human immunodeficiency virus restriction factor BST-2/Tetherin via a {beta}TrCP-dependent mechanism. J Virol. 2009 Aug; 83(16): 7931-47. PMCID: PMC2715753

7: Douglas JL, Whitford JG, Moses AV. Characterization of c-Kit expression and activation in KSHV-infected endothelial cells. Virology. 2009 Aug 1; 390(2): 174-85. PMCID: PMC3489927

8: Douglas JL, Gustin JK, Dezube B, Pantanowitz JL, Moses AV. Kaposi's sarcoma: a model of both malignancy and chronic inflammation. Panminerva Med. 2007 Sep; 49(3): 119-38. Review. PMID: 17912148

9: Douglas JL, Panis ML, Ho E, Lin KY, Krawczyk SH, Grant DM, Cai R, Swaminathan S, Chen X, Cihlar T. Small molecules VP-14637 and JNJ-2408068 inhibit respiratory syncytial virus fusion by similar mechanisms. Antimicrob Agents Chemother. 2005 Jun; 49(6): 2460-6. PMCID: PMC1140497

10: Douglas JL. In search of a small-molecule inhibitor for respiratory syncytial virus. Expert Rev Anti Infect Ther. 2004 Aug; 2(4): 625-39. Review. PMID: 15482225

11: Douglas JL, Panis ML, Ho E, Lin KY, Krawczyk SH, Grant DM, Cai R, Swaminathan S, Cihlar T. Inhibition of respiratory syncytial virus fusion by the small molecule VP-14637 via specific interactions with F protein. J Virol. 2003 May; 77(9): 5054-64. PMCID: PMC153948

12: Douglas JL, Lin WY, Panis ML, Veres G. Efficient human immunodeficiency virus-based vector transduction of unstimulated human mobilized peripheral blood CD34+ cells in the SCID-hu Thy/Liv model of human T cell lymphopoiesis. Hum Gene Ther. 2001 Mar 1; 12(4): 401-13. PMID: 11242532

13: Barrette S, Douglas JL, Seidel NE, Bodine DM. Lentivirus-based vectors transduce mouse hematopoietic stem cells with similar efficiency to Moloney murine leukemia virus-based vectors. Blood. 2000 Nov 15; 96(10): 3385-91. PMID: 11071632

14: Luo T, Douglas JL, Livingston RL, Garcia JV. Infectivity enhancement by HIV-1 Nef is dependent on the pathway of virus entry: implications for HIV-based gene transfer systems. Virology. 1998 Feb 15; 241(2): 224-33. PMID: 9499797

15: O'Neill E, Douglas JL, Chien ML, Garcia JV. Open reading frame 26 of human herpesvirus 8 encodes a tetradecanoyl phorbol acetate- and butyrate-inducible 32-kilodalton protein expressed in a body cavity-based lymphoma cell line. J Virol. 1997 Jun; 71(6): 4791-7. PMCID: PMC191701

16: Chien ML, Foster JL, Douglas JL, Garcia JV. The amphotropic murine leukemia virus receptor gene encodes a 71-kilodalton protein that is induced by phosphate depletion. J Virol. 1997 Jun; 71(6): 4564-70. PMCID: PMC191678

17: Gopalakrishnan S, Douglas JL, Quinlan MP. Immortalization of primary epithelial cells by E1A 12S requires late, second exon-encoded functions in addition to complex formation with pRB and p300. Cell Growth Differ. 1997 May; 8(5): 541-51. PMID: 9149905

18: Douglas JL, Quinlan MP. Structural limitations of the Ad5 E1A 12S nuclear localization signal. Virology. 1996 Jun 15; 220(2): 339-49. PMID: 8661385

19: Douglas JL, Quinlan MP. Efficient nuclear localization and immortalizing ability, two functions dependent on the adenovirus type 5 (Ad5) E1A second exon, are necessary for cotransformation with Ad5 E1B but not with T24ras. J Virol. 1995 Dec; 69(12): 8061-5. PMCID: PMC189754

20: Douglas JL, Quinlan MP. Efficient nuclear localization of the Ad5 E1A 12S protein is necessary for immortalization but not cotransformation of primary epithelial cells. Cell Growth Differ. 1994 May; 5(5): 475-83. PMID: 8049154

21: Quinlan MP, Douglas JL. Immortalization of primary epithelial cells requires first- and second-exon functions of adenovirus type 5 12S. J Virol. 1992 Apr; 66(4): 2020-30. PMCID: PMC288991

22: Douglas JL, Gopalakrishnan S, Quinlan MP. Modulation of transformation of primary epithelial cells by the second exon of the Ad5 E1A12S gene. Oncogene. 1991 Nov; 6(11): 2093-103. PMID: 1945414

OHSU Home | About OHSU | Search | Site Map | Contact OHSU
Health Care Services | Research Programs | Academic & Students | Regional Outreach

OHSU is an equal opportunity, affirmative action institution.
© 2001-2013, Oregon Health & Science University
OHSU Notice of Privacy Practices