Richard Kriwacki, PhD

Richard W. Kriwacki, PhD

Associate Member, St. Jude Faculty
Adjunct Assistant Professor, Molecular Sciences Dept., UT Health Science Center, Memphis

Departments

Structural Biology

Contact Information

Richard Kriwacki, PhD
Structural Biology
MS 311, Room D-1024F
St. Jude Children's Research Hospital
262 Danny Thomas Place
Memphis, TN 38105-3678

Education

MS - University of Connecticut, Storrs, CT
PhD - Yale University, New Haven, CT


Research Interests

My laboratory seeks to understand the molecular basis of regulation of cell division and apoptosis, with special emphasis on the role of tumor suppressor proteins in these vital biological processes.  We apply structural biology and biophysical techniques (NMR spectroscopy, x-ray crystallography, calorimetry, AUC, etc.), as well as biochemical and cell biological methods, to study the details of biomolecular mechanisms from the test tube to cells. Of particular interest is the role of intrinsic protein flexibility in regulatory mechanisms.

p21 and p27 are small proteins that regulate cyclin-dependent kinases (Cdks), the master timekeepers of cell division. p21 and p27 are intrinsically unfolded, or disordered, in isolation and fold upon binding their cyclin/Cdk targets. Past studies have shown that the folding-upon-binding process is sequential and that this mediates specificity for particular Cdk/cyclin complexes. Recently, we discovered that tyrosine kinases (e.g. the leukemogenic kinase, BCR-ABL) directly regulate the inhibitory activity and stability of p27, leading to the concept that p27, and other intrinsically disordered proteins, serve as molecular conduits for signal transduction. In addition to BCR-ABL, a number of other oncogenic tyrosine kinases (e.g. Src and Lyn) have recently been shown to phosphorylate p27 in association with human cancer. Our continuing studies are focused on fully understanding the mechanistic role of the p27 signaling conduit in tumorigenesis.

A new project focuses on regulation of the intrinsic apoptotic pathway by the proteins, p53, BCL-xL and PUMA. Past studies by the Green laboratory at St. Jude have shown that PUMA is unique amongst BH3-only, pro-apoptotic proteins in causing the release of p53 from inactive complexes with BCL-xL.  However, the molecular details of this process have remained a mystery. Using NMR spectroscopy, x-ray crystallography, and a plethora of biochemical and cellular assays, we have discovered a novel molecular mechanism by which PUMA releases p53 from BCL-xL. This mechanism involves drastic, PUMA-induced structural remodeling of BCL-xL.  Continuing studies seek to understand the generality of this mechanism in apoptotic signaling. Further, we seek to understand at the atomic level how binding of PUMA to BCL-xL brings about the gross structural rearrangement that releases p53.  Mutagenesis, calorimetry, biochemical and cellular assays, x-ray crystallography and NMR spectroscopy are being applied to understand the structural, dynamic and thermodynamic features of the PUMA-induced p53 release mechanism.   


Selected Publications

Chipuk JE, Fisher JC, Dillon CP, Kriwacki RW, Kuwana T, Green DR. Mechanism of apoptosis induction by inhibition of the anti-apoptotic BCL-2 proteins. Proc Natl Acad Sci U S A 105:20327 2008. [Dec 12, 2008. Epub]

Teichert A, Arnold LA, Otieno S, Oda Y, Augustinaite I, Geistlinger TR, Kriwacki RW, Guy RK, Bikle DD. Quantification of the vitamin D receptor-coregulator interaction. Biochemistry 48:1454 2009.

Galea CA, Wang Y, Sivakolundu SG, Kriwacki RW. Regulation of cell division by intrinsically unstructured proteins: intrinsic flexibility, modularity, and signaling conduits. Biochemistry 47:7598, 2008.

Galea CA, High AA, Obenauer JC, Mishra A, Park CG, Punta M, Schlessinger A, Ma J, Rost B, Slaughter CA, Kriwacki RW. Large-Scale Analysis of Thermostable, Mammalian Proteins Provides Insights into the Intrinsically Disordered Proteome. J Proteome Res 8:211, 2009. [Dec 9, 2008. Epub ahead of print]

Sivakolundu, SG, Nourse A, Moshiach S, Bothner B, Ashley C, Satumba J, Lahti J, Kriwacki RW. Intrinsically Unstructured Domains of Arf and Hdm2 Form Bi-molecular Oligomeric Structures In Vitro and In Vivo. J Mol Biol 384:240, 2008. Epub 2008 Sep 16.

Galea CA, Nourse A, Wang Y, Sivakolundu SG, Heller WT, Kriwacki RW. Role of intrinsic flexibility in signal transduction mediated by the cell cycle regulator, p27Kip1. J Mol Biol 376:827, 2008. Note: Featured in “News and Views” piece in Nat Chem Biol 4: 229, 2008. 

Kerr ID, Sivakolundu S, Li Z, Buchsbaum JC, Knox LA, Kriwacki R, White SW. Crystallographic and NMR analyses of UvsW and UvsW.1 from bacteriophage T4. J Biol Chem 282:34392, 2007.

Pham TC, Kriwacki RW, Parrill AL. Peptide design and structural characterization of a GPCR loop mimetic. Biopolymers 86:298, 2007.

Zhan J, Easton JB, Huang S, Mishra A, Xiao L, Lacy ER, Kriwacki RW, Houghton PJ. Negative regulation of ASK1 by p21Cip1 involves a small domain that includes Serine 98 that is phosphorylated by ASK1 in vivo. Mol Cell Biol 27:3530, 2007.

Grimmler M, Wang Y, Mund T, Cilensek Z, Keidel EM, Waddell MB, Jakel H, Kullmann M, Kriwacki RW, Hengst L. Cdk-inhibitory activity and stability of p27Kip1 are directly regulated by oncogenic tyrosine kinases. Cell 128:269, 2007.  Note: Featured in “News and Views” piece in Cell 128:241, 2008, and Nat Rev Cancer 7:162, 2007.

Galea CA, Pagala V, Obenauer JC, Park C, Slaughter CA, Kriwacki RW. Proteomic Studies of the Intrinsically Unstructured Mammalian Proteome. J Proteome Res 5:2839, 2006.

Ribeiro RC, Rodriguez-Galindo C, Figueiredo BC, Kriwacki R, Zambetti GP.  Letter to the Editor: Germline TP53 R337H mutation is not sufficient to determine Li-Fraumeni or Li-Fraumeni-like syndrome. Cancer Letters 247:353, 2007.

Jordan JB, Kovacs H, Wang Y, Mobli M, Luo R, Anklin C, Hoch JC, Kriwacki RW. 3D 13C-detected CH3-TOCSY using selectively protonated proteins: Facile methyl resonance assignment and protein structure determination. J Amer Chem Soc 128:9119, 2006.

West AN, Ribeiro RC, Jenkins J, Rodriguez-Galindo C, Figueiredo BC, Kriwacki R, Zambetti GP. Identification of a Novel Germ Line Variant Hotspot Mutant p53-R175L in Pediatric Adrenal Cortical Carcinoma. Cancer Res 66:5056, 2006.

Bowman P, Galea CA, Lacy E, Kriwacki RW. Thermodynamic characterization of interactions between p27(Kip1) and activated and non-activated Cdk2: Intrinsically unstructured proteins as thermodynamic tethers. Biochem Biophys Acta 2006 (Epub ahead of print).

Wang Y, Filippov I, Richter C, Luo R, Kriwacki RW. Solution NMR studies of an intrinsically unstructured protein within a dilute, 75 kDa eukaryotic protein assembly; probing the practical limits for efficiently assigning polypeptide backbone resonances. Chem Biochem 6:2242-2246, 2005.

Galea C, Bowman P, Kriwacki RW. Disruption of an intermonomer salt bridge in the p53 tetramerization domain results in an increased propensity to form amyloid fibrils. Protein Sci 14:2993-3003, 2005.

Sivakolundu SG, Bashford D, Kriwacki RW. Disordered p27Kip1 exhibits intrinsic structure resembling the Cdk2/cyclin A-bound conformation. J Mol Biol 353:1118-1128, 2005.

Luo R, Mann B, Tuomanen E, Kriwacki RW. NMR assignment of the R2 domain of pneumococcal choline binding protein A (CbpA). J Biomol NMR 32:93, 2005.

Lacy ER, Wang Y, Post J, Nourse A, Webb W, Mapelli M, Musacchio A, Siuzdak G, Kriwacki RW. Molecular basis for the specificity of p27 toward cyclin-dependent kinases that regulate cell division. J Mol Biol 349:764-773, 2005.

Luo R, Mann B, Lewis WS, Rowe A, Heath R, Stewart ML, Hamburger AE, Sivakolundu S, Lacy ER, Bjorkman PJ, Tuomanen E, Kriwacki RW. Solution structure of choline binding protein A, the major adhesin of Streptococcus pneumoniae. EMBO J 24:34-43, 2005.

Lacy ER, Filippov I, Lewis WS, Otieno S, Xiao L, Weiss S, Hengst L, Kriwacki RW. P27 binds cyclin-CDK complexes through a sequential mechanism involving binding-induced protein folding. Nat Struct Mol Biol 11:358-364, 2004.

McKenzie PP, Danks MK, Kriwacki RW, Harris LC. P21 Waf1/Cip1 dysfunction in neuroblastoma: a novel mechanism of attenuating G0-G1 cell cycle arrest. Cancer Res 63:3840-3844, 2003.

Lee AS, Galea C, DiGiammarino EL, Jun B, Murti G, Ribeiro RC, Zambetti G, Schultz CP, Kriwacki RW. Reversible amyloid formation by the p53 tetramerization domain and a cancer-associated mutant. J Mol Biol 327:699-709, 2003.

Bothner B, Aubin Y, Kriwacki RW. Peptides derived from two dynamically disordered proteins self-assemble into amyloid-like fibrils. J Amer Chem Soc 125:3200-3201, 2003.

Nishimoto SK, Waite JH, Nishimoto M, Kriwacki RW. Structure, activity, and distribution of fish osteocalcin. J Biol Chem 278:11843-11848, 2003.

Somerville L, Krynetski EY, Krynetskaia NF, Beger RD, Zhang W, Marhefka CA, Evans WE, Kriwacki RW. Structure and dynamics of thioguanine-modified duplex DNA. J Biol Chem 278:1005-1011, 2003.

Jayawardene DS, Liu J, Zhang W, Kriwacki RW. Assignments of the 1H, 13C, and 15N resonances of the winged helix domain of the proto-oncoprotein cQin (FoxG1B). J Biomol NMR 23:243-4, 2002.

DiGiammarino EL, Lee A, Cadwell C, Zhang W, Bothner B, Zambetti G, Ribeiro R, Kriwacki RW. Evidence for a novel molecular mechanism of tumorigenesis: pH-dependent destabilization of the p53 tetramer. Nat Struct Biol 9:12-16, 2002.

Bothner B, Lewis W , DiGiammarino EL, Jason D, Weber JD, Bothner S, Kriwacki RW. Defining the molecular basis of Arf and Hdm2 interactions. J Mol Biol 314:263-277, 2001.

Li N, Zhang W, White SW, Kriwacki RW. Solution structure of the transcriptional activation domain of the bacteriophage T4 protein, MotA. Biochemistry 40:4293-302, 2001.

DiGiammarino EL, Bothner B, Filippov I, Weber JD, Xiao L, Kriwacki RW. Solution structure of the p53 regulatory domain of the ARF tumor suppressor. Biochemistry 40:2379-2386, 2001.

Kriwacki RW, Legge GB, Hommel U, Ramage P, Chung J, Tennant LL, Wright PE, Dyson JH. Assignment of 1H, 13C, and 15N resonances for the I-domain of human leukocyte function associated antigen-1. J Biomol NMR 16:271-272, 2000.

Weber JD, Kuo ML, Bothner B, DiGiammarino EL, Kriwacki RW, Roussel MF, Sherr CJ. Cooperative signals governing ARF-mdm2 interaction and nucleolar localization of the complex. Mol Cell Biol 20:2517-2528, 2000.

Legge GB, Kriwacki RW, Chung J, Hommel U, Ramage P, Case DA, Dyson HJ, Wright PE. NMR solution structure of the inserted domain of human leukocyte function associated antigen-1. J Mol Biol 295:1251-1264, 2000.

Kallen J, Welzenbach K, Ramage P, Geyl D, Kriwacki R, Legge G, Cottens S, Weitz-Schmidt G, Hommel U. Structural basis for LFA-12 inhibition upon lovastatin binding to the CD11a I-domain. J Mol Biol 292:1-9, 1999.

Last update: July 2009

 

Email This Article Email This Article   |   Print Print