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Texas Consortium for Genomic Analysis or TCGA Favorite Outdoor
Sport is Fly Fishing |
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Larry H. Rohde, Ph. D. Associate Professor of Biology and Biotechnology Program Chair for Biotechnology Bayou
Building, Room 3525-6, Box 267 2700 Bay
Area Blvd., Houston, Texas 77058-1098 Office
Phone: 281-283-3743 Fax: 281-283-3709 Is Campus
open or closed because of weather? Contact http://www.uhclemergency.info/ Or call UHCL
Hotline: 281-283-2221 |
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Courses, Independent Studies and
Master Thesis UHCL Biology Website and UHCL Biotechnology Website Current undergraduate and graduate
courses: Genetics, Developmental Biology, Tissue Culture,
Stem Cell Biology Genomics: Sequencing and Analysis Laboratory,
Histology Laboratory Molecular Biology Laboratory. Training of undergraduates and
graduate students in my research laboratory:
I NEED STUDENTS!!!!!!
Fall 2008 and Spring 2009: 1)
NIH Grant
Wrap-up: I need Independent
study students for completing my research on my NIH grant. We will be performing
Co-immunoprecipitation experiments this Fall. Graduate Students need to have
Tissue Culture experience and Molecular Biology Skills. Undergraduate Students need to have courses
in Tissue Culture and Molecular Biology Laboratories. 2)
NSF Grant Continuation: I will need Independent study
students to help create digital Histology library using the Nikon E80i
Microscope. Looking for Biology
students who have extensive computer skills. Courses 2008 to 2009 Courses that
I will be teaching for Fall 2008, Spring 2009 and Summer 2009 semesters
(tentative): Fall: Genetics (Evening Classes two times a
week), Developmental Biology and Tissue Culture Spring: Genetics (Morning Classes two times a
week), Molecular Biology Laboratory and Histology Laboratory Summer: Stem Cell Biology and Genomic Sequencing
Laboratory Future
courses that will be taught: Genes & Development and Eukaryotic Gene Expression PROFESSIONAL MEMBERSHIPS 2003 The
American Society for Biochemistry and Molecular Biology 1998 American
Association for the Advancement of Science Research Description The
p53 tumor suppressor is the most frequent genetic lesion implicated in human
cancers. Mutant forms of p53 are found at frequencies ranging from 70%
of all lung cancers to 20% of all breast cancers (see p53 mutation database
at http://p53.free.fr/).
Although tumor growth in the presence of wild-type p53 may be caused by the
loss of other tumor suppressor genes, the overexpression of p53 inhibitors,
the lack of p53 stimulators, or other possible causes may include factors
that regulate p53 inhibitors and stimulators. Recently, the p53 binding
protein, apoptotic stimulator protein of p53 (ASPP2), was shown to be a major
stimulator of p53-dependent apoptosis and to be down regulated in 23% of
human breast carcinomas (1). In addition, an ASPP family member
called iASPP was shown to be an oncoprotein that inhibits p53 activity
(2). Since these studies suggest that the expression levels of ASPP
family members regulate wild-type p53 activity, then those factors that regulate
the activity of ASPP2 may also influence p53’s apoptotic function during
tumor suppression. Furthermore, the C-terminal fragment of ASPP2
has been shown to interact with several proteins that modulate its role in
regulating apoptosis and p53 activity (3-7). However, all of
these ASPP2 interacting proteins were identified using fragments of these
proteins and not ASPP2 to screen cDNA libraries (3-7). This
presents strong evidence that other proteins inhibit and/or stimulate the
activity of ASPP2. Because of the diversity of these proteins and
their modification of ASPP2's function, these data suggest that other
proteins may also be binding and modulating ASPP2's activity. This
proposal is designed to answer the following questions: What other proteins
are interacting with ASPP2 and how does this interaction influence its
apoptotic function?
Therefore, I hypothesize that ASPP2 interacts with proteins, yet to be
identified, that modulate its stimulatory function for p53-dependent apoptosis. By identifying new ASPP2-binding proteins (ASbps), this
study will expand the current knowledge of ASPP2’s p53-stimulatory function,
identify mechanisms of cancer not due to mutant p53, and possibly elucidate
other ASPP2 regulated pathways. To investigate this hypothesis, I have
proposed the following specific aims: 1. Isolate and identify genes that encode proteins that bind
ASPP2 in the yeast two-hybrid system. In
this specific aim, various domains of ASPP2 will be used in the Matchmaker
yeast two-hybrid system (BD Biosciences Clontech) to screen a placental cDNA
library for putative ASPP2-interacting proteins. Fragments of ASPP2
will be inserted into the DNA-BD vector using standard techniques and a
commercially available placental cDNA-AD vector library will be
screened. Expression of fusion proteins in yeast will be verified and
lack of reporter gene activation by fusion genes will be tested. After
the yeast two-hybrid assay is performed, the putative positives clones will
be analyzed for binding specificity and then sequenced. Putative
positive clones will be tested with an in vitro BD Matchmaker
Co-Immunoprecipitation kit to confirm protein-protein interactions identified
with the yeast two-hybrid assay. 2.
Confirm
binding specificity between ASPP2 and putative ASPP2-binding proteins in
vivo using mammalian cell lines. The
purpose of this specific aim is to eliminate artifacts of the yeast
two-hybrid system by characterizing the protein-protein interaction in mammalian
cell lines. Binding specificity between ASPP2 and ASbps will be
analyzed in vivo using the Matchmaker Mammalian Two-Hybrid system (BD
Biosciences Clontech). This assay will consist of the cotransfection of
plasmids encoding binding partners of Specific Aim 1 and a SEAP reporter gene
into mammalian cell line. The full-length cDNAs of putative positive
clones from the mammalian two-hybrid assay will then be acquired (e.g., ATCC)
and overexpressed in a mammalian cell line containing an inducible ASPP2 protein.
This will be followed by co-immunoprecipitation studies using anti-ASPP2 or
HA-Tag polyclonal antibodies. 3.
Determine
the effects of ASPP2-binding proteins on ASPP2 stimulation of p53’s gene
transactivation or apoptotic induction. Apoptotic and p53-transactivation assays will be
used to characterize ASbps modulation of p53-transactivation via ASPP2.
ASPP2-expressing cell lines over-expressing ASbps will be studied using
standard protocols for apoptosis. p53-transactivation will be
quantitated with a p53-luciferase reporter gene construct cotransfected with
ASbps into ASPP2-inducible cells. Education
Postdoctoral Fellowships Stanford University School of Medicine, Stanford,
CA Department
of Pediatrics 1996 – 1999: Supervised
by Louie Naumovski, M.D., Ph.D. I
studied the function of a p53 binding protein, BBP (or p53BP2), that induces
G2/M cell cycle arrest in cells.
I used the Ecdysone-Inducible Expression system (Invitrogen) to
generate stable clones that will express BBP upon induction with ponasterone
A. Clones were analyzed by FACScan to
determine the affect of BBP expression on cell cycle progression and by using
a luciferase reporter gene linked to p53 response elements to determine if
BBP binds p53 in vivo. Immunofluorescent microscopy was used to colocalized
p53 with wildtype and mutant forms of BBP. Department
of Cardiovascular Medicine 1995 – 1996: Supervised by Richard
Pratt, Ph.D. Laboratory
research focused on the development of smooth muscle cells in the aortic
wall. Rat embryonic and adult,
balloon-injured aortae were collected and cryostat sections were analyzed by
immunohistochemistry for expression of the angiotensin type2 receptor (AT2R)
or for measurement of apoptosis using Hoescht and Apoptag kit methods. I used a C-Imaging System linked to a Leica
microscope to quantitate apoptotic nuclei and AT2R expressing cells in the
tunica media of the aorta. RT-PCR was
used to generate cDNA mimics of each atrial natriuretic peptide (ANP)
receptors, which have an internal deletion of 50 to 100 bp. Competitive RT-PCR on mRNA from each mimic
in combination with total RNA from various samples of embryonic and
balloon-injured aortae was used to quantitate levels of ANP receptor mRNA Ph.D. (Biomedical
Sciences) 1995 in Biochemistry with an emphasis in Developmental Biology
Graduate School of Biomedical Sciences at the University of
Texas Health Science Center, Houston, Texas: Supervised
by Daniel D. Carson, Ph.D. (Dept. of Biochemistry and Molecular
Biology): Currently Chair of the
Department of Biology at the University of Delaware Dissertation: Involvement of Heparin-like Glycosaminoglycans and Expression
of a Novel Heparan Sulfate Binding Proteins (p24) During Human Placentation
and in a Model for Human Implantation Using a
model for human implantation, I identified a heparin/heparan sulfate-interacting
protein (p24 or HIP) that has a role in the initial attachment of embryonic
trophectoderm to uterine epithelial cells.
In the course of this study, I designed, conducted and analyzed
cell-cell and cell-matrix adhesion assays as well as studied binding of
radioligands to cell surface and extracellular matrix components. In addition, I maintained and utilized
H.P.L.C. for analysis of glycoprotein, protein, and glycosaminoglycan samples
using ion exchange and gel-filtration chromatography. Using degenerate oligonucleotide primers, I
conducted reverse transcription-polymerase chain reaction and cloned cDNA
sequences encoding potential heparin binding proteins. I sequenced and performed Northern Blot
analysis, which identified a novel protein, called HIP. I also generated antibodies to HIP,
affinity purified these antibodies, and quantitated antibody production by
E.L.I.S.A. I used Western Blot
analysis to characterize the expression of HIP in cellular fractionation
experiments and utilized indirect immunofluorescent microscopy to localize
HIP on cells and in human uterine-placental tissue from first, second, and
third trimesters. Master of Science Teaching 1986 in Biology
Tarleton State
University, Texas A&M System, Stephenville, Texas: Supervised by Herschel Garner, Ph.D. (Dept. of
Biology): Retired I conducted a capture-recapture study on a population of the Plains
Harvest Mouse, Reithrodontomys montanus (grooved-toothed mouse). I constructed live traps and established a
trapping grid on unused ranch land near Stephenville, TX. Over a 12-month period, traps were rotated,
animals were captured and marked, scat samples were collected and major plant
species were identified.
Bachelor of Science
1984 in Biology
Tarleton State
University, Texas A&M System, Stephenville, Texas My emphasis was in
Field Biology and Geology. Awards
& Scholarships
University of Houston-Clear lake
2007 National
Science Foundation, The Bay Area Houston Partnership for Innovation in Biotechnology
and Life Sciences (PIBLS): Partnerships with San Jacinto Community College
District and University of Texas Medical Branch, (awarded on March 1, 2007)
Funding: $124,227 for three years 2006 ISSO-JSC
joint award with Dr. Honglu Wu (NASA Investigator): Two-year award to fund a Postdoctoral
fellow Position, Title of Award:
Biological Effects of Shielding Parameters across the Bragg Curve of
Energetic Protons and Fe Ions:
Funding: $40,000 per year 2005 Li-COR
Bioscience 2005 Genomics Education Matching Fund Program (awarded on March
15, 2005): Matching Funds ($44,899.50) for the LiCor 4300L with DNA
Sequencing and AFLP Application Package 2004 National
Institute of Health-National Cancer Institute, Academic Research Enhancement
Award R15 (Activated July 1, 2004). Project Title: New Proteins Binding
Apoptotic Stimulator of p53, ASPP2: Funding: $208,513 for three years. Activated: July 1, 2004 2003 Faculty
Research and Support Funds (FRSF), University of Houston-Clear Lake (awarded on December,
2003). (awarded on May,
2003). (awarded on October
23, 2002). (awarded on December 20, 2001). (awarded on July 17, 2000). (awarded on December 1, 1999). (awarded on August 9, 1999) STANFORD
MEDICAL CENTER 1996-1998: An F32
Individual National Research Service Award (NRSA) reference number: 1F32HL09552-01
(HHVJ); National Institute of Health; National Heart, Lung, and Blood Institute. GRADUATE
SCHOOL OF BIOMEDICAL SCIENCE at UTHSC-HOUSTON 1991-1994:
American Legion Auxiliary Predoctoral Fellowship (9/91 to 9/94) PUBLICATIONS Research
Papers: Zhang Y, Rohde,
LH., Emanmi, K., Hammond, D., Casey, R., Mehta, S., Jeevarajan, A.,
Pierson, D. and Wu, H. Suppressed Expression of Non-DSB Repair Genes Inhibits
Gamma-radiation Induced Cytogenetic Repair And Cell Cycle Arrest. DNA
Repair (2008) Accepted for Publication Rohde LH, Ao Y and Naumovski L. p53-Interacting Protein 53BP2 Inhibits
Clonogenic Survival and Sensitizes Cells to Doxorubicin but not
Paclitaxel-induced Apoptosis. Oncogene (2001) 20:2720-2725. Lopez C, Ao Y, Rohde
LH, Perez T., O’Conner D., Lu X., Ford J.M. and Naumovski L. Proapoptotic
p53-Interacting Protein 53BP2 Is Induced by UV Irradiation but Suppressed by
p53. Molecular and Cellular Biology, 2000; 20:8018-8025. Rohde LH, Janatpour MJ, McMaster MT, Fisher SJ, Zhou Y, Lim K-H,
French M, Hoke D. Julian J and Carson DD. Complimentary Expression of
Heparin/Heparan Sulfate Interacting Protein and Perlecan at the Human
Fetal-Maternal Interface. Biol. Reproduction 1998;
58:1075-1083. Rohde LH, Julian J, Babaknia A and Carson DD. Cell surface
expression of HIP, a novel heparin/heparan sulfate binding protein, of human
uterine epithelial cells and cell lines. J. Biol. Chem. 1996;
271:11824-11830. Liu S, Smith SE,
Julian J, Rohde LH, Karin NJ, and Carson DD. cDNA cloning and
expression of HIP, a novel cell surface heparan sulfate/herparin binding
protein of human uterine epithelial cells and cell lines. J. Biol.
Chem. 1996; 271:11817-11823. Rohde LH, and Carson DD. Heparin-like glycosaminoglycans
participate in binding of a human trophoblastic cell line (JAR) to a human
uterine epithelial cell line (RL95). J. Cell. Physiol. 1993;
155:185-196. Raboudi N, Julian J, Rohde
LH, and Carson DD. Identification of cell-surface heparin/heparan
sulfate- binding proteins of a human uterine epithelial cell line
(RL95). J. Biol. Chem. 1992; 267:11930-11939. Book
Chapters: Carson DD, Jacobs AL,
Julian J and Rohde LH. Proteoglycan as modulators of
embryo-uterine interactions. In In Vitro Fertilization and Embryo
Transfer in Primates (Wolf, D.P., Stouffer, R.L., and Brenner, R.M.,
eds) 1993; pp 290-307, Springer-Verlag, New York. Reviews: Carson DD, Rohde
LH, and Surveyor G. Cell surface glycoconjugates as modulators of
embryo attachment to uterine epithelial cells. Int. J. Biochem 1993;
26:1269-1277. Carson
DD, Jacob AJ, Julian J, Rohde LH, and Valdizan MD.
Glycoconjugates as positive and negative modulators of embryo
implantation. Reprod. Fertil. Dev. 1992; 4:271-274. Abstracts: Ye Zhang, Satish Mehta1,
Dianne Hammond1, Duane Pierson, Antony Jeevarajan, Larry Rohde and Honglu Wu, Expression of genes associated with
DNA damage sensing in human fibroblast exposed to low-dose-rate gamma rays.
18th annual NASA Space Radiation Investigators’ Workshop, 2007. Morgado, M.,
McCreight, M., Chuke, C., Hart,C., Moody, T., Laporte, D., Juarez, Y.,
Shaefer, J., Nyholt, K., Spanos, P., and Rohde,
L.H. New Proteins Binding
apoptotic Stimulator of p53. American
Association for the Advancement of Science – Southwest and Rocky Mountain
Division, 82nd Annual Meeting, 2007 Bray, B.A., Grigoreff, Y., Forbus, J.D., Beavers, S.Y., Ruiz, M.I., Butler,
K.A., Warthen, L.M. and Rohde, L.H. Proteins that interact with
the apoptotic stimulator protein of p53, ASPP2, in the yeast two-hybrid
system. American Society for Biochemistry and Molecular Biology Annual
Meeting and 8th IUBMB Conference. 2004 Rohde
LH, Julian J. Fisher SJ and Carson,
DD. Immunological studies of a heparin binding protein from a human
uterine epithelial cell line. Mol. Biol. Cell 1994; 5(suppl):1751 Liu
S, Julian J, Rohde LH and Carson DD. HSBP-1 peptide is a
specific HS/HP-binding peptide. FASEB J. 1994; 8:851. Rohde
LH, Liu S and Carson DD. Cloning of
a heparin-binding protein from a human uterine epithelial cell line.
FASEB J. 1993; 7:1210. Rohde
LH, Raboudi N, and Carson DD.
Proteoglycan involvement in human uterine epithelial cell-trophoblastsic cell
interactions: an In Vitro model of implantation. J. Cell Biol.
1989; 109(suppl):1283. |
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