maryschu@uiuc.edu
161 Edward R. Madigan Laboratory (ERML)
Office: (217) 333-8784
Lab: (217) 333-8785
Fax: (217) 244-1336
Mail to:
Department of Cell and Developmental Biology
University of Illinois at Urbana-Champaign
135 Edward R. Madigan Laboratory (ERML)
1201 W. Gregory Dr.
Urbana, IL 61801
Mary A Schuler
Professor of Cell and Developmental Biology
Professor of Biochemistry
Professor of Plant Biology
NSF2010 site
Education
B.A., 1974 Cornell University (Chemistry and Biology)
Ph.D., 1981 Cornell University (Biochemistry)
Postdoc., 1981-1983 Washington University
Teaching Interests
Molecular biology, biochemistry and genomics: pre-mRNA processing and P450 monooxygenases
One focus of the research in my laboratory is aimed at understanding the factors that influence the removal of introns from pre-mRNAs in plant nuclei and defining the mechanism for intron excision. Although it might be logical to assume that plants and animals share common mechanisms for intron removal, in vivo and in vitro studies have demonstrated that the splicing machineries are not interchangeable between plants and animals or, for that matter, between dicot and monocot plants. Molecular analysis of the intron splice site requirements in plant nuclei using in vivo expression systems has allowed us to demonstrate that plant 5' and 3' splice site recognition depends on having strong AU-transition points between intron and exon that are used for their selection. Current projects are aimed at defining the range of splicing factors mediating constitutive and alternative splicing during plant growth and development. Using Arabidopsis T-DNA knockout/knockout lines and RNA silencing lines depleted for different classes of splicing factors, we are characterizing the role of a number of plant-specific splicing factors as well as general splicing factors. Using microarray genomic approaches, we are analyzing the range of transcripts affected by splicing factor inactivation. Coupled with our current knowledge on required cis-acting sequences, these molecular approaches will eventually identify the trans-acting factors involved in the recognition of plant and other AU-rich introns and–in the long run–help define the optimal sequences needed to express genes/proteins in transgenic plants.
The second major focus in my laboratory is aimed at defining the transcriptional elements regulating expression of plant and insect cytochrome P450 monooxygenase genes. In both plants and insects, these genes encode enzymes critical for synthesis of an array of hormones and defense compounds as well as the metabolism of hydroxylated products such as herbicides and insecticides. Because of this, their transcriptional regulatory cascades respond to natural and synthetic compounds, predators and pathogens in the environment. A variety of RT-PCR and microarray strategies are being used to define the range of plant (Arabidopsis thaliana, Zea mays) P450 genes induced in response to chemical and environmental stresses and the range of insect (Papilio polyxenes, Helicoverpa zea) P450 genes induced in response to plant signaling molecules and toxins. The elements regulating transcription are being identified using promoter:reporter gene fusions (CAT, LUC, GFP) in transgenic plants and in transfected insect cell cultures.
The third focus in my laboratory is aimed at developing molecular models of the catalytic sites in insect P450s metabolizing plant toxins and insecticides and in plant P450s synthesizing an array of secondary products. A variety of protein expression strategies using yeast and baculovirus systems and site-directed changes in catalytic site residues are being used to refine our molecular models and to predict alternate substrates for each of these P450s.
Representative Publications
2008
Ming, R. et al. 2008. The draft genome of the transgenic tropical fruit tree papaya (Carica papaya Linnaeus). Nature 452, 991-996. [Abstract] [Full Article]
Mao, W., A.R. Zangerl, M.R. Berenbaum and M.A. Schuler. 2008b. Metabolism of myristicin by Depressaria pastinacella CYP6AB3v2 and inhibition by its metabolite. Insect Biochem. Mol. Biol. 38, 645-651. [Abstract] [Full Article]
Kühnel, K., N. Ke, M.A. Schuler, S.G. Sligar and I. Schlichting. 2008. Crystal structures of substrate-free and retinoic acid-bound cyanobacterial cytochrome P450 CYP120A1. Biochemistry 47, 6552-6559. [Abstract] [Full Article]
Chiu, T.-L., Z. Wen, S.G. Rupasinghe and M.A. Schuler. 2008. Comparative molecular modeling of Anopheles gambiae CYP6Z1, a mosquito P450 capable of metabolizing DDT. Proc. Natl. Acad. Sci. USA 105, 8855-8860. [Abstract] [Full Article] [PNAS Highlights]
Wang, W., S.G. Rupasinghe, M.A. Schuler and E.G. de Mejia. 2008. Identification of topoisomerase II inhibitory peptides from soy protein hydrolysate by co-immunoprecipitation approach. J. Agric. Food Chem. [Epub ahead of print] [Abstract] [Full Article]
Schuler, M.A. 2008. Splice site requirements and switches in plants. In "Nuclear pre-mRNA processing in plants"; A.S.N. Reddy, M. Golovkin, Eds.; Springer-Verlag.
Mao, W., Berenbaum, M.R., and Schuler, M.A. 2008. Modifications in the N-terminus of an insect cytochrome P450 enhance production of catalytically active protein in baculovirus-Sf9 cell expression systems. Insect Biochem. Mol. Biol., 38:66–75. [Abstract] [Full Article]
2007
Li, X., Schuler, M.A., and Berenbaum, M.R. 2007. Molecular mechanisms of metabolic resistance to synthetic and natural xenobiotics. Annu. Rev. Entomol. 52:231–53. [Abstract] [Full Article]
Li, X., Wen, Z., Bohnert, H.J., Schuler, M.A., and Kushad, M.M. 2007. Myrosinase in horseradish (Armoracia rusticana) root: Isolation of a full-length cDNA and its heterologous expression in Spodoptera frugiperda insect cells. Plant Sci., 172:1095–1102. [Abstract] [Full Article]
Mao, W., Rupasinghe, S., Zangerl, A., Berenbaum, M.R., and Schuler, M.A. 2007. Allelic variation in the Depressaria pastinacella CYP6AB3 protein enhances metabolism of plant allelochemicals by altering a proximal surface residue and potential interactions with cytochrome P450 reductase. J. Biol. Chem., 282:10544–52. [Abstract] [Full Article]
Mao, W., Schuler, M.A., and Berenbaum, M.R. 2007. Cytochrome P450s in Papilio multicaudatus and the transition from oligophagy to polyphagy in the Papilionidae. Insect Mol. Biol., 16:481–490. [Abstract] [Full Article]
Rupasinghe, S., Duan, H., and Schuler, M.A. 2007. Molecular definitions of fatty acid hydroxylases in Arabidopsis thaliana. Proteins, 68: 279–93. [Abstract] [Full Article]
Rupasinghe, S.G., Duan, H., Frericks Schmidt, H.L., Berthold, D., Rienstra, C.M., and Schuler, M.A. 2007. High-yield expression, purification and refolding of isotopically labeled cytochrome P450 monooxygenases for solid-state NMR spectroscopy. Biochim. Biophys. Acta, 1768:3061–70. [Abstract] [Full Article]
Rupasinghe, S., Schuler, M.A., Kagawa, N., Yuan, H., Lei, L., Zhao, B., Kelly, S.L., Waterman, M.R. and Lamb, D.C. 2007. The cytochrome P450 gene family CYP157 does not contain EXXR in the K-helix reducing the absolute conserved P450 residues to a single cysteine. FEBS Lett., 580:6338–6342. [Abstract] [Full Article]
Rupasinghe, S.G., Wen, Z., Chiu, T.L. and Schuler, M.A. 2007. Helicoverpa zea CYP6B8 and CYP321A1: different molecular solutions to the problem of metabolizing plant toxins and insecticides. Protein Eng. Des. Sel., 20:615–24. [Abstract] [Full Article]
Schuler, M.A. and Sligar, S.G. 2007. Diversities and similarities in P450 systems: an introduction. In "The ubiquitous roles of cytochrome P450 proteins", Vol. 3 of 'Metal Ions in Life Sciences'; A. Sigel, H. Sigel, R. K. O. Sigel, Eds.; John Wiley & Sons, Ltd., Chichester, UK. Pp. 1-26.
Zeng, R.S., Wen, Z., Niu, G., Schuler, M.A., and Berenbaum, M.R. 2007. Allelochemical induction of cytochrome P450 monooxygenases and amelioration of xenobiotic toxicity in Helicoverpa zea. J. Chem. Ecol., 33:449–461. [Abstract] [Full Article]
2006
Baudry, J., Rupasinghe, S., and Schuler, M.A. 2006. Class-dependent sequence alignment strategy improves the structural and functional modeling of P450s. Protein Eng. Des. Sel., 19:345–53. [Abstract] [Full Article]
Claudianos, C., Ranson, H., Johnson, R.M., Biswas, S., Schuler, M.A., Berenbaum, M.R., Feyereisen, R., and Oakeshott, J.G. 2006. A deficit of detoxification enzymes: pesticide sensitivity and environmental response in the honeybee. Insect Mol. Biol., 15:615–36. [Abstract] [Full Article]
Duan, H. and Schuler, M.A. 2006. Heterologous expression and strategies for encapsulation of membrane-localized plant P450s. Phytochemistry Reviews, 5:507–23. [Abstract] [Full Article]
Honey Bee Genome Sequencing Consortium. 2006. Insights into social insects from the genome of the honey bee Apis mellifera. Nature, 443:931–49. [Abstract] [Full Article]
Johnson, R.M., Wen, Z., Schuler, M.A., and Berenbaum, M.R. 2006. Mediation of pyrethroid insecticide toxicity to honey bees (Hymenoptera: Apidae) by cytochrome P450 monooxygenases. J. Econ. Entomol. 99:1046–50. [Abstract]
Mao, W., Rupasinghe, S., Zangerl, A., Schuler, M.A., and Berenbaum, M.R. 2006. Remarkable substrate-specificity of CYP6AB3 in Depressaria Pastinacella, a highly specialized caterpillar. Insect Mol. Biol., 15:169–79. [Abstract] [Full Article]
Rupasinghe, S., and Schuler, M.A. 2006. Homology modeling of plant P450s. Phytochemistry Reviews, 5:473–505. [Abstract] [Full Article]
Schuler, M.A., Duan, H., Bilgin, M., and Ali, S. 2006. Arabidopsis P450s through the looking glass: a window on plant biochemistry. Phytochemistry Reviews, 5:205–37. [Abstract] [Full Article]
Wen, Z., Berenbaum, M.R., and Schuler, M.A. 2006. Inhibition of CYP6B1-mediated detoxification of xanthotoxin by plant allelochemicals in the black swallowtail (Papilio polyxenes). J. Chem. Ecol., 32:507–22. [Abstract] [Full Article]
Wen, Z., Rupasinghe, S., Niu, G., Berenbaum, M.R., and Schuler, M.A. 2006. CYP6B1 and CYP6B3 of the black swallowtail (Papilio polyxenes): adaptive evolution through subfunctionalization. Mol. Biol. Evol., 23:2434–43. [Abstract] [Full Article]
Zeng, R., Niu, G., Wen, Z., Schuler, M.A., and Berenbaum, M.R. 2006. Toxicity of aflatoxin B1 to helicoverpa zea and bioactivation by cytochrome P450 monooxygenases. J. Chem. Ecol., 32:1459–71. [Abstract] [Full Article]
2005
Brown, R.P., McDonnell, C.M., Berenbaum, M.R., and Schuler, M.A. 2005. Regulation of an insect cytochrome P450 monooxygenase gene (CYP6B1) by Aryl Gydrocarbon and Xanthotoxin response cascades. Gene, 358:39–52. [Abstract] [Full Article]
Duan, H., Huang, M.-Y., Palacio, K., and Schuler, M.A. 2005. Variations in CYP74B2 (Hydroperoxide Lyase) gene expression differentially affect hexenal signaling in the columbia and landsberg erecta ecotypes of Arabidopsis. Plant Physiol, 139:1529–44. [Abstract] [Full Article]
Duan, H. and Schuler, M.A. 2005. Differential expression and evolution of Arabidopsis CYP86A subfamily. Plant Physiol, 137:1067–81. [Abstract] [Full Article]
Ke, N., Baudry, J., Makris, T.M., Schuler, M.A., and S.G. Sligar. 2005. A retinoic acid binding cytochrome P450: CYP120A1 from Synechocystis sp. PCC 6803. Arch. Biochem. Biophys, 436:110-20. [Abstract] [Full Article]
Thimmapuram, J., Duan, H., Liu, L., and Schuler, M.A. 2005. Bicistronic and fused monocistronic transcripts are derived from adjacent loci in the Arabidopsis Genome. RNA 11, 128–38. [Abstract] [Full Article]
Wen, Z., Baudry, J., Berenbaum, M.R., and Schuler, M.A. 2005. Ile115Leu mutation in the SRS1 region of an insect cytochrome P450 (CYP6B1) compromises substrate turnover via changes in a predicted product release channel. Prot. Eng. Design Select., 18:191–9. [Abstract] [Full Article]