PHD, Biological Chemistry, University of California at Los Angeles, 1973
B.A., Chemistry (Conc in BIochemistry), San Jose State University, 1968
Interdisciplinary and Emerging Signature Programs
Cardiovascular
Research Interests
<P><B>CENTRAL HYPOTHESIS - </B>We hypothesize that in accordance with the strict temporal and spatial control of NO production in vascular endothelial cells, argininosuccinate synthase, the rate-limiting step of the citrulline-NO cycle, is coordinately and tightly regulated via translational and posttranslational mechanisms, in addition to transcriptional regulation. This premise underlies the rationale for the specific aims our present research which focuses on posttranscriptional events that regulate AS function. First, we hypothesize that the determinants of AS subcellular localization are AS-interacting proteins which guide AS to distinct destinations within the cell. Methodology used to examine this hypothesis employs: 1) immunofluorescence microscopy; 2) pull-down analysis; 3) immunoprecipitation analysis; 4) tandem affinity purification analysis; 5) mass-spectrometry; and 6) NO determinations. Second, we hypothesize that a small protein encoded by an upstream open reading frame (uORF) represses AS mRNA translation under stress conditions by routing stalled AS mRNA particles into discrete cytoplasmic foci known as stress granules. To test this hypothesis we employ: 1) in vitro translation; 2) RNA immunoprecipitation; 3) RNA EMSA; 4) polysome profiling; 5) immunofluorescence microscopy; 6) western blotting; and 7) fluorescence in situ hybridization (FISH). Third, we hypothesize that endothelial AS protein is phosphorylated at serine/theronine residues which mediate an acute response, altering AS activity. To test this hypothesis the following methodology has been employed: 1) proteomics and mass-spectrometry; 2) mutational analysis; 3) NO determinations; 4) AS activity assays; and 5) western blotting. Fourth, to translate our cell culture work to a whole animal system, we are studying how changes in AS expression correlate with impaired vascular endothelial NO production in a diabetic rat model system. Experimental approaches include: 1) animal model characterization; 2) real time RT-PCR; 3) western blotting; 4) NO measurements; 5) immunofluorescence microscopy; and 6) FISH. Overall, results from this research have the potential to distinguish new therapeutic targets to be used to prevent or inhibit vascular endothelial dysfunction found in diabetes, obesity and other related vascular diseases.</P>
Awards/Honors
Medical Education Award, Department of Molecular Medicine (USF College of Medicine, Tampa, FL - 2007)
Silver Performance Award for Outstanding Contributions 2006/2007 (USF College of Medicine, Tampa, FL - 2007)
Honorary Member of the Golden Key International Honor Society (University of South Florida, Tampa, FL - 2006)
Outstanding Contribution to the Program in Undergraduate Medical Education (USF College of Medicine, Tampa, FL - 2003)
Presidents Award for Faculty Excellence (USF College of Medicine, Tampa, FL - 2003)
Outstanding Contribution to the Program of Undergraduate Medical Education (USF College of Medicine, Tampa, FL - 2001)
Professorial Excellence Program Award, 1998-1999. (USF College of Medicine, Tampa, FL - 1999)
Distinguished Teacher Award (1996), presented from the Medical Class of 1996. (USF College of Medicine, Tampa, FL - 1996)
Outstanding Advisor Award from the Association of Medical Science Graduate Students (USF College of Medicine, Tampa, FL - 1991)
The Jerome Krivanek Distinguished Teacher Award (University of South Florida, Tampa, FL - 1991)
Outstanding Pre-Clinical Professor (USF College of Medicine, Tampa, FL - 1990)
Fellow of the Jane Coffin Childs Memorial Fund for Medical Research (Stanford University, Standord, Calif - 1972)
Margaret B. Kraatz Cancer Award for Outstanding Graduate Student Research (UCLA, Los Angeles, Calif - 1972)
Graduated with Great Distinction (San Jose State University, San Jose, California - 1968)
Memberships
Member (American Heart Association, 2007 - Present)
Member (North American Association for the Study of Obesity, 2007 - Present)
Member (The RNA Society, 2007 - Present)
Member (American Society of Biological Chemists, 2007 - Present)
Member (H. Lee Moffitt Cancer Center & Research Institute, 2007 - Present)
Member (American Society of Microbiology, 2007 - Present)
Member (Institute for Biomolecular Science, 2007 - Present)
Recent Publications
Flam BR, Eichler DC, Solomonson LP. Endothelial nitric oxide production is tightly coupled to the citrulline-NO cycle.Nitric oxide : biology and chemistry / official journal of the Nitric Oxide Society. 17(3-4) : 115-21
Bansal V, Rodriguez P, Wu G, Eichler DC, Zabaleta J, Taheri F, Ochoa JB. Citrulline can preserve proliferation and prevent the loss of CD3 zeta chain under conditions of low arginine.JPEN. Journal of parenteral and enteral nutrition. 28(6) : 423-30
Haines RJ, Pendleton LC, Eichler DC. Argininosuccinate synthase: at the center of arginine metabolism.International journal of biochemistry and molecular biology. 2(1) : 8-23, 2011.
Goodwin BL, Corbin KD, Pendleton LC, Levy MM, Solomonson LP, Eichler DC. Troglitazone up-regulates vascular endothelial argininosuccinate synthase.Biochemical and biophysical research communications. 370(2) : 254-8, 2008.
Goodwin BL, Pendleton LC, Levy MM, Solomonson LP, Eichler DC. Tumor necrosis factor-alpha reduces argininosuccinate synthase expression and nitric oxide production in aortic endothelial cells.American journal of physiology. Heart and circulatory physiology. 293(2) : H1115-21, 2007.
Pendleton LC, Goodwin BL, Solomonson LP, Eichler DC. Regulation of endothelial argininosuccinate synthase expression and NO production by an upstream open reading frame.The Journal of biological chemistry. 280(25) : 24252-60, 2005.
Goodwin BL, Solomonson LP, Eichler DC. Argininosuccinate synthase expression is required to maintain nitric oxide production and cell viability in aortic endothelial cells.The Journal of biological chemistry. 279(18) : 18353-60, 2004.
Solomonson LP, Flam BR, Pendleton LC, Goodwin BL, Eichler DC. The caveolar nitric oxide synthase/arginine regeneration system for NO production in endothelial cells.The Journal of experimental biology. 206(Pt 12) : 2083-7, 2003.
Patel NA, Eichler DC, Chappell DS, Illingworth PA, Chalfant CE, Yamamoto M, Dean NM, Wyatt JR, Mebert K, Watson JE, Cooper DR. The protein kinase C beta II exon confers mRNA instability in the presence of high glucose concentrations.The Journal of biological chemistry. 278(2) : 1149-57, 2003.
Diamond FB, Eichler DC. Leptin and the adipocyte endocrine system.Critical reviews in clinical laboratory sciences. 39(4-5) : 499-525, 2002.
Pendleton LC, Goodwin BL, Flam BR, Solomonson LP, Eichler DC. Endothelial argininosuccinate synthase mRNA 5''-untranslated region diversity. Infrastructure for tissue-specific expression.The Journal of biological chemistry. 277(28) : 25363-9, 2002.
Flam BR, Hartmann PJ, Harrell-Booth M, Solomonson LP, Eichler DC. Caveolar localization of arginine regeneration enzymes, argininosuccinate synthase, and lyase, with endothelial nitric oxide synthase.Nitric oxide : biology and chemistry / official journal of the Nitric Oxide Society. 5(2) : 187-97, 2001.
Patel NA, Chalfant CE, Watson JE, Wyatt JR, Dean NM, Eichler DC, Cooper DR. Insulin regulates alternative splicing of protein kinase C beta II through a phosphatidylinositol 3-kinase-dependent pathway involving the nuclear serine/arginine-rich splicing factor, SRp40, in skeletal muscle cells.The Journal of biological chemistry. 276(25) : 22648-54, 2001.
Hellermann GR, Flam BR, Eichler DC, Solomonson LP. Stimulation of receptor-mediated nitric oxide production by vanadate.Arteriosclerosis, thrombosis, and vascular biology. 20(9) : 2045-50, 2000.
Diamond FB, Eichler DC. Leptin: molecular biology, physiology, and relevance to pediatric practice.Advances in pediatrics. 46: 151-87, 1999.
Eichler DC, Root AW, Duckett G, Moore KL, Diamond FB. A spun-column assay for determination of leptin binding in serum.Analytical biochemistry. 267(1) : 100-3, 1999.
Patel NA, Chalfant CE, Yamamoto M, Watson JE, Eichler DC, Cooper DR. Acute hyperglycemia regulates transcription and posttranscriptional stability of PKCbetaII mRNA in vascular smooth muscle cells.The FASEB journal : official publication of the Federation of American Societies for Experimental Biology. 13(1) : 103-13, 1999.
Chalfant CE, Watson JE, Bisnauth LD, Kang JB, Patel N, Obeid LM, Eichler DC, Cooper DR. Insulin regulates protein kinase CbetaII expression through enhanced exon inclusion in L6 skeletal muscle cells. A novel mechanism of insulin- and insulin-like growth factor-i-induced 5'' splice site selection.The Journal of biological chemistry. 273(2) : 910-6, 1998.
Diamond FB, Eichler DC, Duckett G, Jorgensen EV, Shulman D, Root AW. Demonstration of a leptin binding factor in human serum.Biochemical and biophysical research communications. 233(3) : 818-22, 1997.
Eichler DC, Craig N. Processing of eukaryotic ribosomal RNA.Progress in nucleic acid research and molecular biology. 49: 197-239, 1994.
Eichler DC. Characterization of a nucleolar 2''-O-methyltransferase and its involvement in the methylation of mouse precursor ribosomal RNA.Biochimie. 76(12) : 1115-22, 1994.
Eichler DC. Information processing and gene expression.The Journal of the Florida Medical Association. 80(1) : 63-6, 1993.
Eichler DC, Liberatore JA, Shumard CM. Selection of a preribosomal RNA processing site by a nucleolar endoribonuclease involves formation of a stable complex.Nucleic acids research. 21(24) : 5775-81, 1993.
Segal DM, Eichler DC. A nucleolar 2''-O-methyltransferase. Specificity and evidence for its role in the methylation of mouse 28 S precursor ribosomal RNA.The Journal of biological chemistry. 266(36) : 24385-9, 1991.
Shumard CM, Torres C, Eichler DC. In vitro processing at the 3''-terminal region of pre-18S rRNA by a nucleolar endoribonuclease.Molecular and cellular biology. 10(8) : 3868-72, 1990.
Segal DM, Eichler DC. The specificity of interaction between S-adenosyl-L-methionine and a nucleolar 2''-O-methyltransferase.Archives of biochemistry and biophysics. 275(2) : 334-43, 1989.
Shumard CM, Eichler DC. Ribosomal RNA processing. Limited cleavages of mouse preribosomal RNA by a nucleolar endoribonuclease include the early +650 processing site.The Journal of biological chemistry. 263(36) : 19346-52, 1988.
Eichler DC, Eales SJ. Specificity of a nucleolar 2''-O-methyltransferase for RNA substrates.Biochemical and biophysical research communications. 155(1) : 530-7, 1988.
Eichler DC, Raber NK, Shumard CM, Eales SJ. Isolation and characterization of a nucleolar 2''-O-methyltransferase from Ehrlich ascites tumor cells.Biochemistry. 26(6) : 1639-44, 1987.
Eichler DC, Solomonson LP, Barber MJ, McCreery MJ, Ness GC. Radiation inactivation analysis of enzymes. Effect of free radical scavengers on apparent target sizes.The Journal of biological chemistry. 262(20) : 9433-6, 1987.
Ness GC, Sample CE, Smith M, Pendleton LC, Eichler DC. Characteristics of rat liver microsomal 3-hydroxy-3-methylglutaryl-coenzyme A reductase.The Biochemical journal. 233(1) : 167-72, 1986.
Barber MJ, Solomonson LP, Eichler DC. Spin-labeled erythrocyte membranes: direct identification of nitroxide-conjugated proteins.Biochemical and biophysical research communications. 127(3) : 793-8, 1985.
Eichler DC, Barber MJ, Solomonson LP. Anti-nitroxide immunoglobulin G: analysis of antibody specificity and their application as probes for spin-labeled proteins.Biochemistry. 24(5) : 1181-6, 1985.
Eichler DC, Eales SJ. Purification and properties of a novel nucleolar exoribonuclease capable of degrading both single-stranded and double-stranded RNA.Biochemistry. 24(3) : 686-91, 1985.
Lasater LS, Eichler DC. Isolation and properties of a single-strand 5''----3'' exoribonuclease from Ehrlich ascites tumor cell nucleoli.Biochemistry. 23(19) : 4367-73, 1984.
Eichler DC, Eales SJ. The effect of RNA secondary structure on the action of a nucleolar endoribonuclease.The Journal of biological chemistry. 258(16) : 10049-53, 1983.
Eichler DC, Eales SJ. Isolation and characterization of a single-stranded specific endoribonuclease from Ehrlich cell nucleoli.The Journal of biological chemistry. 257(23) : 14384-9, 1982.
Saporta S, Eichler DC. Conjugated dimers of horseradish peroxidase as neuroanatomical tracers.Brain research. 250(1) : 149-53, 1982.
Eichler DC, Tatar TF, Lasater LS. Effect of human placental ribonuclease inhibitor in cell-free ribosomal RNA synthesis.Biochemical and biophysical research communications. 101(2) : 396-403, 1981.
Eichler DC, Tatar TF. Properties of a purified nucleolar ribonuclease from Ehrlich ascites carcinoma cells.Biochemistry. 19(13) : 3016-22, 1980.
Eichler DC, Lehman IR. On the role of ATP in phosphodiester bond hydrolysis catalyzed by the recBC deoxyribonuclease of Escherichia coli.The Journal of biological chemistry. 252(2) : 499-503, 1977.
Wang TS, Eichler DC, Korn D. Effect of Mn2+ on the in vitro activity of human deoxyribonucleic acid polymerase beta.Biochemistry. 16(22) : 4927-34, 1977.
Eichler DC, Wang TS, Clayton DA, Korn D. In vitro replication of mitochondrial DNA. Elongation of the endogenous primer sequence in D loop mitochondrial DNA by human DNA polymerase beta.The Journal of biological chemistry. 252(21) : 7888-93, 1977.
Eichler DC, Fisher PA, Korn D. Effect of calcium on the recovery and distribution of DNA polymerase alpha from cultured human cells.The Journal of biological chemistry. 252(11) : 4011-4, 1977.
Uyemura D, Eichler DC, Lehman IR. Biochemical characterization of mutant forms of DNA polymerase I from Escherichia coli. II. The polAex1 mutation.The Journal of biological chemistry. 251(13) : 4085-9, 1976.
Glitz DG, Eichler DC, Angel L. On the use of polyacrylamide gel electrophoresis in the analysis of the action of free and insolubilized nucleases on bacteriophage RNA.Analytical biochemistry. 62(2) : 552-67, 1974.
Glitz DG, Eichler DC. Characterization of a guanylic acid specific ribonuclease from Aspergillus fumigatus.Biochemistry. 11(10) : 1746-54, 1972.
