**CURRICULUM VITAE**

**PATRICIA MARTINE DAY**

**PERSONAL**

Place of Birth: London, England

Citizenship: USA

**ADDRESS**

Work: Room 4112/Building 37

LCO/CCR

NCI/NIH

9000 Rockville Pike

Bethesda, MD 20892

301/496-6539

301/480-5322 (fax)

**EDUCATION**

September 1979 - May 1983. B.S. Biology Major

St. Lawrence University

Canton, New York 13617

September 1985 - November 1991. Ph.D.

Department of Microbiology and Immunology

University of North Carolina at Chapel Hill

Chapel Hill, North Carolina 27599

Advisor: Dr. Jeffrey A. Frelinger

Thesis: Analysis of a Nonclassical Class I Molecule, Qb-1

**HONORS AND AWARDS**

September 1979 - May 1983. New York State Regents Scholarship

September 1985 - November 1991. Graduate Assistantship

September 1985 - November 1991. Graduate Tuition and Fee Scholarship

September 1985 - November 1991. Anne Rauch Graduate Scholarship

September 2005. Federal Technology Transfer Award

**EMPLOYMENT**

November 1991 - June 1996. IRTA Postdoctoral Research Fellow

Viral Immunology Section

Laboratory of Viral Diseases

National Institute of Allergy and Infectious Diseases

National Institutes of Health

Bethesda, Maryland 20892.

My work in this laboratory was focused on the cell biology of MHC class
I restricted antigen processing. By confocal microscopy and flow
cytometry I examined the intracellular trafficking of class I molecules.
One project resulted in the demonstration that peptide-receptive
molecules, those that lack stably associated antigenic peptide, are
transported to the cell surface with greater efficiency in cells that
contain the peptide transporter, TAP, and those that are TAP-deficient.
This suggests that TAP functions to increase the number of
peptide-receptive molecules at the cell surface. A second project
involved the transport of extracellular antigenic peptides into the
endoplasmic reticulum. These peptides associate with the appropriate
class I molecule via a TAP-independent pathway. This finding
demonstrates the existence of a pathway, likely vesicular in nature,
that conveys small extracellular substances to the ER without traversing
the Golgi complex or the cytosol.

July 1996 -- June 1999 Biotechnology Fellow

Laboratory of Cellular Oncology

Department of Basic Sciences

National Cancer Institute

National Institutes of Health

Bethesda, Maryland 20892

June 1999 -- present Staff Scientist

Laboratory of Cellular Oncology

Department of Basic Sciences

National Cancer Institute

National Institutes of Health

Bethesda, Maryland 20892

My major projects in LCO have involved applying cell biological
techniques to questions of papillomavirus entry and early events in
infection.

In one of my early studies in this area I defined the cellular pathway
utilized by papillomaviruses for uptake and intracellular transport
during the establishment of infection.

In a separate project, I developed an assay to evaluate viral uncoating
by assessing antibody accessibility to the carboxyl terminus of L2 and
the viral genome. This allowed me to determine that the minor capsid
protein, L2, mediates the delivery of the genome to a transcriptionally
active nuclear domain, ND10. I have found that intact ND10 are also
critical for efficient establishment of infection.

I also demonstrated that the L2 capsid protein is modified by the
cellular protease, furin during endocytosis. This cleavage is critical
for the infectious process. In the absence of furin cleavage both the L2
protein and the genome are retained within the endosomal compartment.
This was the first identification of a role for furin in the entry of
any virus.

More recently I have evaluated the mechanism of neutralization of
various monoclonal antibodies. This led to the understanding of which
different regions of the capsid interact with either cell surface
receptors or the extracellular matrix. I have also been evaluating
conformational changes of the viral capsid that occur on the cell
surface. I made a panel of monoclonal antibodies that recognize an open
capsid conformation that occurs soon after cell

binding. We are hopeful that these studies will lead to the development
of a cross-neutralizing immunogen.

In addition to these projects within LCO, I have established
collaborative projects with other NIH laboratories and also extramural
laboratories. Currently I am collaborating with Ari Helenius at the
Swiss Federal Institute of Technology in Zurich, to further investigate
the endocytic pathway with a specific emphasis on cellular signaling
molecules that are necessary for infection. Another current
collaborative effort, with Richard Roden at Johns Hopkins University,
involves the investigation of the mechanism of neutralization of anti-L2
monoclonal antibodies.

**TEACHING**

September 1994-June 1997. Instructor

Introductory Microbiology lecture and laboratory course

\[organized and taught all lectures and laboratories\]

Montgomery College

Rockville, Maryland

May, June 1992. Virology/Immunology Lectures

NIH Program for Advanced High School Students

\[general lectures and laboratory demonstration\]

January 1988 - May 1988. Graduate Teaching Assistant

September 1986 - December 1986. Microbiology Laboratory

University of North Carolina at Chapel Hill.

\[included lecture on theory in addition to supervising

laboratory experiments\]

January 1988 - December 1990. Immunology tutor

\[individual tutoring to medical and dental students\]

University of North Carolina at Chapel Hill.

**MENTORING**

May 1994 - August 1994. Supervisor of NIH Summer Intern Student

\[directed and supervised laboratory project\]

June 2004- May 2005 Mentor

Oversaw project for postbaccalaureate research fellow

Rebecca Richards

We described the requirement for furin cleavage of the minor capsid
protein during endocytosis of virions. This cleavage was found to be a
highly conserved activity in all papillomaviruses. Mature PV capsids
need to undergo a conformational change after ECM or cell surface
binding to render them susceptible to furin cleavage. Prevention of this
cleavage event blocks infection.

June 2005- June 2006 Mentor

Oversaw project for postbaccalaureate research fellow

Sarah Galanti

We constructed a transposon-insertion based mutant library of the HPV16
L2 gene. This library yielded over 100 L2 mutants that result in \<10%
of wild-type infectivity. The mutants are being evaluated for
interaction with L1 and the pseudogenome, cell binding and entry
phenotypes, endosomal lysis activity, and colocalization with ND10
during assembly and infection.

July 2006- July 2007 Mentor

Oversaw project for postbaccalaureate research fellow

Michelle Mergler

We successfully made monoclonal antibodies that specifically recognize
an open conformation of the HPV16 capsid. The immunogen appears to mimic
the conformational changes in the capsid that occurs after ECM and/or
cell surface binding. We are currently evaluating the utility of these
antibodies for microscopic analyses and virus neutralization studies.

July 2007- present Mentor

Overseeing project for postbaccalaureate research fellow

Katherine Johnson

Our laboratory's recent development of the cervicovaginal challenge
model for PV infection enables us to conduct a wide variety of *in vivo*
studies. We are currently initiating a project to look at the pathway of
infection within the *in vivo* system.

**PUBLICATIONS**

**1. Day, P.M.** and J. A. Frelinger. *Differential expression of Q4
proteins (Qb-1) in fibroblasts and lymphocytes.* (1991) J. Immunol.
147:3427-3433.

**2.** Bennink, J.R., R. Anderson, I Bacik, J. Cox, **P. Day**, Y. Deng,
C. Lapham, H. Link, G. Russ, and J.W. Yewdell. *Antigen processing:
where tumor-specific T-cell responses begin.* (1993) J. Immunotherapy.
14:202-208.

**3. Day, P.M.**, F. Esquivel, J. Lucsko, J.R. Bennink, and J.W.
Yewdell. *Effect of the transporter associated with antigen processing
(TAP) on the generation and intracellular trafficking of peptide
receptive major histocompatibility complex class I molecules* (1995)
Immunity. 2:137-147.

**4. Day, P.M.**, J. W. Yewdell, A. Porgador, R.N. Germain, and J.R.
Bennink. *Direct delivery of* *exogenous MHC class I molecule-binding
oligopeptides to the endoplasmic reticulum of viable cells*. (1997)
Proc. Natl. Acad. Sci. USA 94:8064-8069.

**5. Day, P.M.**, R.B.S. Roden, D.R. Lowy, and J.T. Schiller. *The
Papillomavirus Minor Capsid Protein, L2, Induces Localization of the
Major Capsid Protein, L1, and the Viral Transcription/Replication
Protein, E2, to PML Oncogenic Domains.* (1998) J. Virol. 72(1):142-150.

**6.** Anton, L.C., U. Schubert, I. Bacik, M.F. Princiotta, P.A.
Wearsch, J. Gibbs, **P.M. Day**, C. Realini, M.C. Rechsteiner, J.R.
Bennink, and J.W. Yewdell. *Intracellular Localization of Proteasomal
Degradation of a Viral Antigen.* (1999) J. Cell Biol. 146(1): 113-124.

**7.** Fedele, M., G. Benvenuto, R. Pero, B. Majello, S. Battista, F.
Lembo, E. Vollono, **P.M. Day**, M. Santoro, L. Lania, C.B. Bruni, A.
Fusco, and L. Chiariotti. *A novel member of the BTB/POZ Family, PATZ,
associates with the RNF4 RING finger protein and acts as a
transcriptional repressor.* (2000) J. Biol. Chem. 275(11): 7894-7901

**8.** Okun, M.M., **P.M. Day**, H.L. Greenstone, F.P. Booy, D.R. Lowy,
J.T. Schiller, and R.B.S. Roden. *L1 Interaction Domains of
Papillomavirus L2 Necessary for Viral Genome Encapsidation.* (2001) J.
Virol. 75(9):4322-4342.

**9.** Lenz, P., **P.M. Day**, Y-Y Pang, S. Frye, P.N. Jensen, D.R.
Lowy, and J.T. Schiller. *Papillomavirus-like particles induce acute
activation of dendritic cells*. (2001) J. Immunol. May
1;166(9):5346-5355.

**10.** Roden, R.B., **P.M. Day**, B.K. Bronzo, W.H. Yutzy 4th, Y. Yang
, D.R. Lowy, J.T. Schiller. *Positively charged termini of the L2 minor
capsid protein are necessary for papillomavirus infection.* (2001) J.
Virol. Nov;75(21):10493-10497.

**11.** Young, P.J., **P.M. Day**, J. Zhou, E.J. Androphy, G.E. Morris,
C.L. Lorson. *A direct interaction between the survival motor neuron
protein and p53 and its relationship to spinal muscular atrophy*. (2002)
J. Biol. Chem. Jan 25;277(4):2852-2859.

**12.** Lenz, P., C.D. Thompson, **P.M. Day**, S.M. Bacot, D.R. Lowy,
and J.T. Schiller. *Interaction of papillomavirus virus-like particles
with human myeloid antigen-presenting cells.* (2003) Clin. Immunol.
106(3):231-237.

**13. Day, P.M**., D.R. Lowy, and J.T. Schiller. *Papillomaviruses
infect cells via a clathrin- dependent pathway.* (2003) Virology. Mar
1;307(1):1-11.

This manuscript characterized the viral entry pathway. We co-localized
internalized virus with cellular markers of the clathrin endocytic
pathway and also inhibited infection with a panel of biochemical
inhibitors with a profile matching clathrin-dependent uptake.

**14.** Yang, R., **P.M. Day**, PM, W.H. Yutzy 4^th^, K.Y. Lin, C.F.
Hung, and R.B. Roden. Cell surface-binding motifs of L2 that facilitate
papillomavirus infection. (2003) J. Virol. Mar;77(6):3531-3541.

**15.** [**Day, P.M.**, C.C. Baker, D.R. Lowy, and J.T.
Schiller.](http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15383670&query_hl=1)
*Establishment of papillomavirus infection is enhanced by promyelocytic
leukemia protein (PML) expression.* (2004) Proc Natl Acad Sci USA. Sep
28;101(39):14252-14257.

This manuscript identified a role for PML in the enhancement of viral
infectivity in the early part of the life cycle. It also introduced a
method to evaluate viral uncoating microscopically. Confocal microscopic
analyses showed the accumulation of both L2 and viral DNA at distinct
nuclear domains identified as nuclear domain 10 (ND10). This
localization was proven critical for the efficient establishment of
infection. We proposed a model in which L2 chaperones the viral genome
to ND10 to efficiently initiate viral transcription.

**16. Day, P.M.** and J.T. Schiller. Early Events in the Papillomaviral
Lifecycle. p 175-192., in Papillomavirus Research: From Natural History
to Vaccines and Beyond. Caister Academic Press 2005.

This book chapter reviewed the early events in papillomavirus binding
and entry and events leading to the initial establishment of
papillomavirus infection.

**17.** Kaemper, N., **P.M. Day**, T. Nowak, H.-C. Selinka, L. Florin,
J. Bolscher, L. Hilbig, J.T. Schiller, M. Sapp. *A membrane
destablilizing peptide in capsid protein L2 required for egress of
papillomavirus genomes from endosomes.* (2006) J. Virol.
Jan;80(2):759-768.

This paper demonstrated that the C-terminus of L2 functions to disrupt
endocytic vesicles during the infectious process, allowing escape of the
L2/genome complex into the cytosol.

**18.** Buck, C.B., **P.M.** [**Day,** C.D. Thompson, J. Lubkowski, W.
Lu, D.R. Lowy, and J.T.
Schiller.](http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15383670&query_hl=1)
*Human Alpha Defensins Block Papillomavirus Infection.* (2006) Proc Natl
Acad Sci USA. Jan 31;103(5):1516-1521.

**19.** Richards, R.R., D.R. Lowy, J.T. Schiller, and **P.M. Day**.
*Cleavage of the papillomavirus minor capsid protein, L2, at a furin
consensus site is necessary for infection.* (2006) Proc Natl Acad Sci
USA. Jan 31;103(5):1522-1527.

This manuscript demonstrated that the L2 capsid protein is modified by
the cellular protease, furin during the entry process. This cleavage is
critical for the infectious process. This was the first identification
of a role for furin during the endocytosis of any virus.

**20.** [**Day, P.M.**, C.D. Thompson, C.B. Buck, Y.Y. Pang, D.R. Lowy,
and J.T.
Schiller.](http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15383670&query_hl=1)
*Neutralization of human papillomavirus with monoclonal antibodies
reveals different mechanisms of inhibition.* (2007) J. Virol.
Aug;81(16):8784-8792.

This study characterized the mechanism of neutralization of several
neutralizing monoclonal antibodies. Antibodies with epitopes on the apex
of the capsomers did not affect cell surface association, but
dramatically inhibited binding to the ECM. However, the cell bound virus
was inefficiently internalized. In contrast, an antibody with a binding
site on the side wall of the pentamers, blocked cell surface binding,
resulting in sequestration of the capsids on the ECM. Our results
suggest that ECM binding involves the apex of the capsomers, while HSPG
binding involves the clefts between the capsomers.

21\. Baege, A.C., **P.M. Day**, B. Taylor, J. Vogel, R. Schlegel, and
C.C. Baker. *Putative human uterine cervical stem cells display
increased binding of papillomavirus-like particles.* (2007) submitted to
Proc Natl Acad Sci U S A.

22\. **Day,P.M.**, R. Gambhira, R.B.S. Roden, D. R. Lowy, and J. T.
Schiller. *Conformational changes of the HPV16 capsid on the cell
surface revealed by furin cleavage and neutralization by anti-L2
antibodies.* manuscript in preparation.

Our earlier work led us to believe that furin cleavage of the N-terminus
of L2 exposes a binding site on L2 for a critical cellular factor.
Consistent with this conjecture, the major cross-neutralizing epitopes
of L2 also map to this region. We have found that furin cleavage on the
cell surface results in exposure of this cross-neutralizing epitope.
This implies that papillomaviruses have evolved to hide this critical
region from the immune system. We also have gained insight into how to
how to construct potentially cross-neutralizing immunogens.

**INVITED SPEAKING APPEARANCES**

UCLA Symposia on Molecular and Cellular Biology, 1989

Steamboat Springs, CO

NIH Immunology Seminar Series, 1994

Bethesda, MD

Ninth International Congress of Immunology, 1995

San Francisco, CA

Mount St. Mary's College, 1995

Emmitsburg, MD

Niagara University, 1996

Niagara, NY

Cornell College, 1996

Mount Vernon, IA

Sixteenth International Papillomavirus Conference, 1997

Siena, Italy

NIH Virology Interest Group, 1998

Bethesda, MD

NIH Research Festival: Nuclear Structure and Function Symposium, 1998

Bethesda, MD

NIH Virology Interest Group, 2001

Bethesda, MD

EMBO Workshop: The Structural Biology of Small DNA Tumor Viruses, 2001

Siena, Italy

Nineteenth International Papillomavirus Conference, 2001

Florianopolis, Brazil

Twenty-first International Papillomavirus Conference, 2004

Mexico City, Mexico

Twenty-second International Papillomavirus Conference, 2005

Vancouver, British Columbia

NIH Virology Interest Group, 2005

Bethesda, MD

Ninth International Conference on Malignancies in AIDS, 2005

Bethesda, MD

Louisiana State University, Shreveport, LA

Feist-Weiller Cancer Center, Spring Seminar Series, 2006

Johns Hopkins University, Baltimore, MD

Department of Pathology Seminar, 2006

University of North Carolina at Chapel Hill, Chapel Hill, NC

Department of Microbiology and Immunology Seminar Series, 2006

Twenty-third International Papillomavirus Conference, 2006

Prague, Czech Republic

Fourth International Workshop on the Structural Biology of Small DNA
Tumor Viruses, 2007

Siena, Italy

**ADDITIONAL**

Ad hoc reviewer for Journal of General Virology

Ad hoc reviewer for Journal of Virology