Thomas Jefferson University - John F. Klement
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John F. Klement
Dermatology and Cutaneous Biology
Thomas Jefferson University
Jefferson Medical College
Primary Appointment:
Department of Dermatology & Cutaneous Biology
Adjunct Appointment:
Department of Microbiology and Immunology
Rank:
Research Associate Professor
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Mailing Address
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BLSB 412: 233 S. 10th Street
Philadelphia, Pennsylvania 19107
United States
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Qualifications
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2007-Present: Research Associate Professor of Dermatology and Cutaneous Biology and Microbiology and Immunology. TJU
1995-2007: Research Assistant Professor of Dermatology and Microbiology and Immunology; Director of the Dermatology Transgenic Mouse Core. TJU
1991-1995: Postdoctoral position with Dr. Colin L. Stewart and Dr. Craig A. Rosen. Roche Institute of Molecular Biology , Nutley, NJ
1986-1991: Postdoctoral position with Dr. Joram Piatigorsky. NEI, NIH, Bethesda, MD
1986: Ph.D. Microbiology, Rutgers University & Rutgers Medical School; Department of Microbiology, University of Medicine and Dentistry of New Jersey (Currently Robert Wood Johnson Medical School); Piscataway, NJ
1982 M.S. Microbiology, Rutgers University, NJ
1980: B.S. Graduated with Honors in Biology from Moravian College, Bethlehem, PA
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Expertise and Research Interests
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Major Interest: Gene Regulation
Past Research at TJU:
My initial goal at TJU was to establish the Dermatology Transgenic Mouse Core to generate animal models of dermatological diseases. The animal models were constructed by gene ablation (knock-out) using mouse ES cells and gene-targeting technology. The Dermatology Transgenic Mouse Core also generated mice by pronuclear microinjection of DNA.
Models Developed:
1) dystrophic epidermolysis bullosa by deletion of the Col7a1 gene encoding type VII collagen.
2) junctional epidermolysis bullosa by deletion of LamC2 gene encoding laminin-gamma-2 (with Leena Pulkkinen)
3) pseudoxanthoma elasticum by deletion of Abcc6 gene encoding multi-drug-resistance protein 6 (Mrp6)(with Leena Pulkkinen).
Other gene ablations include genes encoding ladinin (unpublished), periplakin, and several genes under industry contract. I also collaborated with other investigators on their genes of interest (see publications).
Current Interests:
The Dermatology Transgenic Mouse Core ceased operations in 2005 due to termination of core funding and conclusion of most of the industrial contracts. Currently, I collaborate with Dr. Jouni Uitto on Abcc6 and Samd9L genes in mice. SamD9L is closely related to human SAMD9 gene that is linked to development of familial tumorial calcinosis.
With the reduction of the Core, I am focusing on my own research interests in the field of gene expression. I am currently studying protein-protein interactions with Mrp6, SamD9, and SamD9L and am hoping to branch out into a more independent research direction.
Expertise:
My expertise ranges from DNA cloning, generation of cell lines, gene promoter studies, and generation and characterization of transgenic mice. My graduate work focused on elucidating the interactions of the bacteriophage RNA polymerases from T3 and T7 with their cognate promoters. This work involved over-expression of the polymerases in bacteria and their subsequent purification, generation of mutant promoter constructs, and in vitro transcription assays. In my post-doctoral work I switched to eukaryotic systems (chicken lens, mice) to study gene expression in the ocular lens (NEI) and cloning genes, and generating gene ablations, in proteins involved in the NF-kappaB signal transduction pathway (RIMB). Both of my post-doctoral posts involved cell culture, promoter assays, transgenic mice, DNA cloning, protein-DNA binding assays, western, northern, and southern blots, basic histology and immunohistochemistry/fluorescence, etc. Currently, I am learning to use the yeast-2-hybrid system to find protein-protein interactions, focusing initially on interactions with human and mouse Mrp6, human SAMD9, and human and mouse SAMD9L.
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Other Expertise
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Teaching:
Participate in 2 courses.
1) Bacterial Genetics section of the graduate-level General Microbiology course (Dr. Kishore Alugupalli, director) with the Department of Microbiology and Immunology.
2) Generation and Societal Impact of Transgenic Animals of the master''s course on Research Animals (Dr. Judy Daviau, director)
Research Animal Use Regulations -- IACUC:
I have also served on the Institutional Animal Care and Use Committee (IACUC) from about 1997 to the present and served as chair for 2006 and 2007. I am familiar with research animal regulations regarding mice and other animals.
Service:
Served on several NIH study sections and departmental and university committees.
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Industrial Relevance
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The industrial relevance of my research is significant. Pharmaceutical companies can utilize animal models of human disease in efforts to develop effective therapeutics. Gene expression, protein-protein interactions, etc., study the basic pathomechanism of disease. By elucidation of these cellular signal transduction pathways, and other protein-protein interactions, it is possible to design or screen for drugs that would repress or enhance gene expression resulting in amelioration of disease.
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Keywords
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Dermatology; gene expression; transgenesis; transgenic mice; gene-targeting; microinjection; protein-protein interactions; gene regulation; promoter; DNA binding; transcription; membrane proteins; signal transduction; microbiology
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Publications
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- 1: Katsuta Y, Ogura Y, Iriyama S, Goetinck PF, Klement JF, Uitto J, Amano S.; Fibulin-5 accelerates elastic fibre assembly in human skin fibroblasts. Exp Dermatol. 2008 Mar 12. PMID: 18341572
- 2: Sevilla LM, Nachat R, Groot KR, Klement JF, Uitto J, Djian P, Määttä A, Watt, FM. Mice deficient in involucrin, envoplakin, and periplakin have a defective epidermal barrier. J Cell Biol. 2007 Dec 31;179(7):1599-612. PMID: 18166659
- 3: Sicot FX, Tsuda T, Markova D, Klement JF, Arita M, Zhang RZ, Pan TC, Mecham RP, Birk DE, Chu ML. Fibulin-2 is dispensable for mouse development and elastic fiber formation. Mol Cell Biol. 2008 Feb;28(3):1061-7. Epub 2007 Dec 10. PMID: 18070922
- 4: Li Q, Jiang Q, Larusso J, Klement JF, Sartorelli AC, Belinsky MG, Kruh GD, Uitto J. Targeted ablation of Abcc1 or Abcc3 in Abcc6(-/-) mice does not modify the ectopic mineralization process. Exp Dermatol. 2007 Oct;16(10):853-9. PMID: 17845218
- 5: Klement JF, Matsuzaki Y, Jiang QJ, Terlizzi J, Choi HY, Fujimoto N, Li K, Pulkkinen L, Birk DE, Sundberg JP, Uitto J. Targeted ablation of the Abcc6 gene results in ectopic mineralization of connective tissues. Mol Cell Biol. 2005 Sep;25(18):8299-310. PMID: 16135817
- 6: Aho S, Li K, Ryoo Y, McGee C, Ishida-Yamamoto A, Uitto J, Klement JF. Periplakin gene targeting reveals a constituent of the cornified cell envelope dispensable for normal mouse development. Mol Cell Biol. 2004 Jul;24(14):6410-8. PMID: 15226441
- 7: Naso M, Uitto J, Klement JF. Transcriptional control of the mouse Col7a1 gene in keratinocytes: basal and transforming growth factor-beta regulated expression. J Invest Dermatol. 2003 Dec;121(6):1469-78. PMID: 14675198
- 8: Meng X, Klement JF, Leperi DA, Birk DE, Sasaki T, Timpl R, Uitto J, Pulkkinen L. Targeted inactivation of murine laminin gamma2-chain gene recapitulates human junctional epidermolysis bullosa. J Invest Dermatol. 2003 Oct;121(4):720-31. PMID: 14632187
- 9: Colombo M, Brittingham RJ, Klement JF, Majsterek I, Birk DE, Uitto J, Fertala A. Procollagen VII self-assembly depends on site-specific interactions and is promoted by cleavage of the NC2 domain with procollagen C-proteinase. Biochemistry. 2003 Oct 7;42(39):11434-42. PMID: 14516194
- 10: Heinonen S, Männikkö M, Klement JF, Whitaker-Menezes D, Murphy GF, Uitto J. Targeted inactivation of the type VII collagen gene (Col7a1) in mice results in severe blistering phenotype: a model for recessive dystrophic epidermolysis bullosa. J Cell Sci. 1999 Nov;112 ( Pt 21):3641-8. PMID: 10523500
- 11: Kouba DJ, Chung KY, Nishiyama T, Vindevoghel L, Kon A, Klement JF, Uitto J, Mauviel A. Nuclear factor-kappa B mediates TNF-alpha inhibitory effect on alpha 2(I)collagen (COL1A2) gene transcription in human dermal fibroblasts. J Immunol. 1999 Apr 1;162(7):4226-34. PMID: 10201951
- 12: Klement JF, Rice NR, Car BD, Abbondanzo SJ, Powers GD, Bhatt PH, Chen CH, Rosen CA, Stewart CL. IkappaBalpha deficiency results in a sustained NF-kappaB response and severe widespread dermatitis in mice. Mol Cell Biol. 1996 May;16(5):2341-9. PMID: 8628301
- 13: Klement JF, Cvekl A, Piatigorsky J. Functional elements DE2A, DE2B, and DE1A and the TATA box are required for activity of the chicken alpha A-crystallin gene in transfected lens epithelial cells. J Biol Chem. 1993 Mar 25;268(9):6777-84. PMID: 8454650
- 14: Donovan DM, Sax CM, Klement JF, Li X, Chepelinsky AB, Piatigorsky J. Conservation of mouse alpha A-crystallin promoter activity in chicken lens epithelial cells. J Mol Evol. 1992 Oct;35(4):337-45. PMID: 1404419
- 15: Ruben SM, Klement JF, Coleman TA, Maher M, Chen CH, Rosen CA. I-Rel: a novel rel-related protein that inhibits NF-kappa B transcriptional activity. Genes Dev. 1992 May;6(5):745-60. PMID: 1577270
- 16: Narayanan R, Klement JF, Ruben SM, Higgins KA, Rosen CA. Identification of a naturally occurring transforming variant of the p65 subunit of NF-kappa B. Science. 1992 Apr 17;256(5055):367-70. PMID: 1566083
- 17: Alemany J, Klement JF, Borrás T, De Pablo F. DNA binding factors which interact with the Sp1 site of the chicken delta 1-crystallin promoter are developmentally regulated. Biochem Biophys Res Commun. 1992 Mar 16;183(2):659-65. PMID: 1550573
- 18: Ruben SM, Narayanan R, Klement JF, Chen CH, Rosen CA. Functional characterization of the NF-kappa B p65 transcriptional activator and an alternatively spliced derivative. Mol Cell Biol. 1992 Feb;12(2):444-54. PMID: 1732726
- 19: Chepelinsky AB, Piatigorsky J, Pisano MM, Dubin RA, Wistow G, Limjoco TI, Klement JF, Jaworski CJ. Lens protein gene expression: alpha-crystallins and MIP. Lens Eye Toxic Res. 1991;8(2-3):319-44. Review. PMID: 1911643
- 20: Sax CM, Klement JF, Piatigorsky J. Species-specific lens activation of the thymidine kinase promoter by a single copy of the mouse alpha A-CRYBP1 site and loss of tissue specificity by multimerization. Mol Cell Biol. 1990 Dec;10(12):6813-6. PMID: 2247086
- 21: Klement JF, Moorefield MB, Jorgensen E, Brown JE, Risman S, McAllister WT. Discrimination between bacteriophage T3 and T7 promoters by the T3 and T7 RNA polymerases depends primarily upon a three base-pair region located 10 to 12 base-pairs upstream from the start site. J Mol Biol. 1990 Sep 5;215(1):21-9. PMID: 2204706
- 22: Klement JF, Wawrousek EF, Piatigorsky J. Tissue-specific expression of the chicken alpha A-crystallin gene in cultured lens epithelia and transgenic mice. J Biol Chem. 1989 Nov 25;264(33):19837-44. PMID: 2584197
- 23: Ling ML, Risman SS, Klement JF, McGraw N, McAllister WT. Abortive initiation by bacteriophage T3 and T7 RNA polymerases under conditions of limiting substrate. Nucleic Acids Res. 1989 Feb 25;17(4):1605-18. Erratum in: Nucleic Acids Res 1989 Jun 12;17(11):4430. PMID: 2646596
- 24: Brown JE, Klement JF, McAllister WT. Sequences of three promoters for the bacteriophage SP6 RNA polymerase. Nucleic Acids Res. 1986 Apr 25;14(8):3521-6. PMID: 3010240
- 25: Morris CE, Klement JF, McAllister WT. Cloning and expression of the bacteriophage T3 RNA polymerase gene. Gene. 1986;41(2-3):193-200. PMID: 3011596
- 26: Bailey JN, Klement JF, McAllister WT. Relationship between promoter structure and template specificities exhibited by the bacteriophage T3 and T7 RNA polymerases. Proc Natl Acad Sci U S A. 1983 May;80(10):2814-8. PMID: 6574450
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Individual Expertise profile of
John F. Klement, Copyright © John F. Klement.
Last Updated
by John Klement : Thursday, August 21, 2008 11:00:49 AM
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