 Perry J. BlackshearAdjunct Professor of Biochemistry, Consulting Professor of Medicine, and Director of Clinical Research, NIEHSStudies in polypeptide hormone interaction. Contact InformationTelephone: (919) 541-4899 Fax: (919) 541-4571 e-mail black009@niehs.nih.gov Building 101 NIEHS 111 Alexander Drive RTP NC 27709 Clinical Res NIEHS PO Box 12233 RTP, NC 27709 Education
Research InterestsThe major subject of study in our laboratory is in the general area of regulation of gene expression by modulation of mRNA decay. Rates of mRNA decay can be altered by a variety of environmental and physiological influences, through trans-acting RNA binding proteins, as well as through intrinsic RNA stability and instability motifs. Our lab studies a small family of four proteins in the mouse that represent a novel class of CCCH tandem zinc finger RNA binding proteins. The prototype of this protein family, tristetraprolin or TTP, is rapidly and dramatically induced in cells, at the transcriptional level, in response to hormones and other stimuli. Using the “knockout mouse” technique, we found that TTP is an RNA binding protein that acts in normal physiology to regulate the secretion of the clinically important cytokines tumor necrosis factor alpha (TNF) and granulocyte-macrophage colony-stimulating factor (GM-CSF). TTP acts by destabilizing these mRNAs, by a process that involves the initial stimulation of deadenylation, or removal of the polyA tail. Gene knockouts of the other known proteins in this family have also been made recently in the laboratory, demonstrating roles for the other family members in normal formation of the fetal circulation and in hematopoiesis, and we are currently evaluating each phenotype to determine the roles of the other family members in normal mRNA metabolism. Single family members are expressed in other organisms, such as Drosophila and S. pombe, and these are being studied as well because of the relative genetic tractability of these model organisms. Ongoing studies are focusing on the regulation of the biosynthesis of the CCCH zinc finger proteins; regulation of their nucleocytoplasmic shuttling; regulation of their phosphorylation and its effects on RNA binding activity; and effects of binding proteins. We are also interested in determining the possible relevance of these proteins to human autoimmune diseases such as rheumatoid arthritis and Crohn’s disease, and to the effects of environmental influences such as bacterial endotoxins. Recent Publications1. Lai, W.S., Carballo, E., Thorn, J.M., Kennington, E.A. and Blackshear, P.J. (2000) Interactions of CCCH zinc finger proteins with mRNA. 1. Binding of tristetraprolin-related zinc finger proteins to AU-rich elements and destabilization of mRNA. J. Biol. Chem., 275:17827-19837. More… 2. Carballo, E., Lai, W.S. and Blackshear, P.J. (2000) Evidence that tristetraprolin (TTP) is a physiological regulator of granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA deadenylation and stability. Blood 95:1891-1899. More… 3. Carballo, E, Pitterle, DM, Stumpo, DJ, Sperling, RT and Blackshear, PJ. (1999) Phagocytic and/or macropinocytic activity in MARCKS-deficient macrophages and fibroblasts. Am. J. Physiol. 277:C163-C173. More… 4. Lai, WS, Carballo,E, Strum, JS, Kennington, EA, Phillips, RS and Blackshear, PJ. (1999) Evidence that tristetraprolin (TTP) binds to AU-rich elements and promotes the deadenylation and destabilization of tumor necrosis factor a mRNA. Mol. Cell. Biol. 19:4311-4323. More… 5. De, J, Lai, WS, Thorn, JM, Goldsworthy, SM, Liu, X, Blackwell, TK and Blackshear, PJ. (1999) Identification of four CCCH zinc finger proteins in Xenopus, including a novel vertebrate protein with four zinc fingers and severely restricted expression. Gene 228: 133-145. More… 6. Lai WS, Thompson MJ and Blackshear PJ. (1998) Characteristics of the intron involvement in the mitogen-induced expression of Zfp-36. J. Biol. Chem. 273:506-517. More… 7. Kim HS, Swierczynski SL, Tuttle JS, Lai WS and Blackshear PJ. (1998) Transgenic complementation ofMARCKS deficiency with a non-myristoylatable, pseudo-phosphorylated form of MARCKS: Evidence for simultaneous positive and dominant-negative effects on central nervous system development. Devel. Biol. 200:146-157. More… 8. Stumpo DJ, Eddy RL, Jr., Haley LL, Sait S, Shows TB, Lai WS, Young WS III, Speer MC, Dehijia A, Polymeropoulos M and Blackshear PJ. (1998) Promoter sequence, expression and fine chromosomal mapping of the human gene (MLP) encoding the MARCKS-like protein: Identification of neighboring and linked polymorphic loci for MLP and MACS and use in the evaluation of human neural tube defects. Genomics 49:253-264. More… 9. Carballo E, Lai, WS and Blackshear PJ. (1998) Feedback inhibition of macrophage tumor necrosis factor a production by tristetraprolin. Science 281:1001-1005. More… 10. McNamara, RK, Stumpo, DJ, Morel, LM, Lewis, MH, Wakeland, EK, Blackshear, PJ and Lenox, RH. (1998) Effect of reduced myristoylated alanine-rich C kinase substrate expression on hippocampal mossy fiber development and spatial learning in mutant mice: Transgenic rescue and interactions with gene background. Proc. Natl. Acad. Sci. USA 95: 14517-14522. More… 11. McNamara, RK, Stumpo, DJ, Morel, LM, Lewis, MH, Wakeland, EK, Blackshear, PJ and Lenox, RH. (1998) Effect of reduced myristoylated alanine-rich C kinase substrate expression on hippocampal mossy fiber development and spatial learning in mutant mice: Transgenic rescue and interactions with gene background. Proc. Natl. Acad. Sci. USA 95: 14517-14522. More… 12. Carballo, E, Gilkeson, GS and Blackshear, PJ (1997). Bone marrow transplantation reproduces the tristetraprolin-deficiency syndrome in recombination activating gene-2 ( 13. Spizz, G and Blackshear, PJ (1997). Identification and characterization of cathepsin B as the cellular MARCKS cleaving enzyme. J. Biol. Chem., 272: 23784-23842. More… 14. Shi Y, Sullivan SK, Pitterle DM, Kennington EA, Graff JM, Blackshear PJ. (1997) Mechanismsof MARCKS gene activation during Xenopus development. J. Biol. Chem. 272: 29290-29300. More… 15. Taylor, GA, Carballo, E, Lee, DM, Lai, WS, Thompson, MJ, Patel, DD, Schenkman, DI, Gilkeson, GS, Broxmeyer, HE, Haynes, BF and Blackshear, PJ (1996). A pathogenetic role for TNF alpha in the syndrome of cachexia, arthritis, and autoimmunity resulting from tristetraprolin (TTP) deficiency. Immunity 4:445-454. More… 16. Swierczynski, SL and Blackshear, PJ (1996) Myristoylation-dependent and electrostatic interactions exert independent effects on the membrane association of the myristoylated alanine-rich protein kinase C substrate protein in intact cells. J. Biol. Chem. 271:23424-23430. More… 17. Blackshear, PJ, Lai, WS, Tuttle, JS, Stumpo, DJ, Kennington, E, Nairn, AC and Sulik, KK (1996). Developmental expression of MARCKS and protein kinase C in mice in relation to the exencephaly resulting from MARCKS deficiency. Devel. Brain Res. 96:62-75. More… 18. Swierczynski, SL, Siddhanti, SR, Tuttle, JS and Blackshear, PJ (1996) Nonmyristoylated MARCKS complements some but not all of the developmental defects associated with MARCKS deficiency in mice Devel. Biol. 179:135-147. More… |
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