Maria Antonietta Ragusa
e-mail: marag AT unipa.it
affiliation: Dip. STEMBIO
research area(s): Molecular Biology, Developmental Biology
Course:
Cell Biology
University/Istitution: Università di Palermo
University/Istitution: Università di Palermo
Degrees.
1992 : Degree in Biological Science, Palermo University, with a mark of 110/110 with honours. Title of the thesis: beta-tubulin expression in P. lividus sea urchin embryo.
1997 : Ph.D. in Cellular and Developmental Biology, Palermo University. Title of the thesis: alfa and beta tubulin expression during sea urchin embryogenesis. Isolation of alfa-2-tubulin gene.
1998-2000 –Postdoctoral scholarship in “Area della Biologia”.
1999: “Embryology: Concepts and Techniques in Modern Developmental Biology” course at Marine Biological Laboratory, Woods Hole, Massachussetts (USA)
Position.
Since January 3rd, 2005: Researcher in Molecular Biology at the Cellular and Developmental Biology Department of Palermo University.
Research Interests.
Molecular and developmental biology.
Transcription regulation of alpha and beta tubulin genes during sea urchin development. Mitochondrial chaperonin HSP60 in sea urchin embryos. HSP40 involvement in sea urchin ciliogenesis and in different stresses. P38 MAPK role on embryo skeletogenesis. Molecular response to heavy metal stress.
Other Interests.
Beta-2-glycoprotein I (b2GPI or ApolipoproteinH) expression in cultured human cells and in rat tissues.
Bioinformatics.
Techniques.
Nucleic acid extraction and analysis, PCR, RT-PCR, qPCR.
Southern, Northern and Western blot analyses.
Cloning and sequencing.
Generation and screening of genomic and cDNA libraries.
Primer extension analysis.
EMSA. Footprinting.
Microinjection in sea urchin eggs.
Culture of sea urchin embryos under physiological or stressing conditions.
In situ hybridization: whole-mount and paraffin sections.
Immunohistochemistry.
Educational Activity.
2000/2001: Recombinant DNA technology,
2001/2002: Molecular biology,
2002/2003 & 2003/2004: Molecular biology, Biological macromolecules,
2004/2005 & 2005/2006: Molecular biology lab., Biological macromolecules, Molecular biology,
2006/2007, 2007/2008, 2008/2009: Molecular biology II, Nucleic acid structure and function
2009/2010: Molecular biology II, Recombinant DNA technology and Bioinformatics application
2010/2011: Recombinant DNA technology and Bioinformatics application
1992 : Degree in Biological Science, Palermo University, with a mark of 110/110 with honours. Title of the thesis: beta-tubulin expression in P. lividus sea urchin embryo.
1997 : Ph.D. in Cellular and Developmental Biology, Palermo University. Title of the thesis: alfa and beta tubulin expression during sea urchin embryogenesis. Isolation of alfa-2-tubulin gene.
1998-2000 –Postdoctoral scholarship in “Area della Biologia”.
1999: “Embryology: Concepts and Techniques in Modern Developmental Biology” course at Marine Biological Laboratory, Woods Hole, Massachussetts (USA)
Position.
Since January 3rd, 2005: Researcher in Molecular Biology at the Cellular and Developmental Biology Department of Palermo University.
Research Interests.
Molecular and developmental biology.
Transcription regulation of alpha and beta tubulin genes during sea urchin development. Mitochondrial chaperonin HSP60 in sea urchin embryos. HSP40 involvement in sea urchin ciliogenesis and in different stresses. P38 MAPK role on embryo skeletogenesis. Molecular response to heavy metal stress.
Other Interests.
Beta-2-glycoprotein I (b2GPI or ApolipoproteinH) expression in cultured human cells and in rat tissues.
Bioinformatics.
Techniques.
Nucleic acid extraction and analysis, PCR, RT-PCR, qPCR.
Southern, Northern and Western blot analyses.
Cloning and sequencing.
Generation and screening of genomic and cDNA libraries.
Primer extension analysis.
EMSA. Footprinting.
Microinjection in sea urchin eggs.
Culture of sea urchin embryos under physiological or stressing conditions.
In situ hybridization: whole-mount and paraffin sections.
Immunohistochemistry.
Educational Activity.
2000/2001: Recombinant DNA technology,
2001/2002: Molecular biology,
2002/2003 & 2003/2004: Molecular biology, Biological macromolecules,
2004/2005 & 2005/2006: Molecular biology lab., Biological macromolecules, Molecular biology,
2006/2007, 2007/2008, 2008/2009: Molecular biology II, Nucleic acid structure and function
2009/2010: Molecular biology II, Recombinant DNA technology and Bioinformatics application
2010/2011: Recombinant DNA technology and Bioinformatics application
1. Gene transcriptional regulation in sea urchin.
Alpha- and beta-tubulin are fundamental components of the eukaryotic cytoskeleton and cell division machinery. Tubulin expression is carefully controlled and most eukaryotes express multiple tubulin genes in specific regulatory or developmental contexts. The PlTalpha2 is a sea urchin P.lividus gene coding for a neural isotype of alpha tubulin. In fact, alpha2 transcript is expressed starting from blastula stage and it is localized to the neural cells that at pluteus stage constitute the ciliary band and the ganglia.
We study the tubulin gene (in particular PlTalpha2) expression regulation utilizing many different approaches, both biochemical (EMSA, footprinting) and functional (gene transfer experiments), to identify cis –acting elements. We perform also DNaseI hypersensitivity studies and Chromatin Immunoprecipitation assays to look at the promoter chromatin structure.
Moreover, to identify gene regulatory elements, we use a combination of bioinformatic approaches such as comparative genomics and phylogenetic footprinting.
2. Molecular biology approaches to understanding stress response mechanisms
Heavy metals have many toxic effects on marine organisms. In particular sea urchin embryos grown in 1 mM cadmium cloride fail to develop normally. We study the early defense strategies activated by P. lividus 30 hours phenotypically normal embryos, in response to exposition to lower doses of cadmium, analyzing the induced transcriptome and comparing it to that of control embryos by RDA technique and subsequent quantitative real-time RT-PCR. Bioinformatics tools are frequently utilized to analyze the results.
Alpha- and beta-tubulin are fundamental components of the eukaryotic cytoskeleton and cell division machinery. Tubulin expression is carefully controlled and most eukaryotes express multiple tubulin genes in specific regulatory or developmental contexts. The PlTalpha2 is a sea urchin P.lividus gene coding for a neural isotype of alpha tubulin. In fact, alpha2 transcript is expressed starting from blastula stage and it is localized to the neural cells that at pluteus stage constitute the ciliary band and the ganglia.
We study the tubulin gene (in particular PlTalpha2) expression regulation utilizing many different approaches, both biochemical (EMSA, footprinting) and functional (gene transfer experiments), to identify cis –acting elements. We perform also DNaseI hypersensitivity studies and Chromatin Immunoprecipitation assays to look at the promoter chromatin structure.
Moreover, to identify gene regulatory elements, we use a combination of bioinformatic approaches such as comparative genomics and phylogenetic footprinting.
2. Molecular biology approaches to understanding stress response mechanisms
Heavy metals have many toxic effects on marine organisms. In particular sea urchin embryos grown in 1 mM cadmium cloride fail to develop normally. We study the early defense strategies activated by P. lividus 30 hours phenotypically normal embryos, in response to exposition to lower doses of cadmium, analyzing the induced transcriptome and comparing it to that of control embryos by RDA technique and subsequent quantitative real-time RT-PCR. Bioinformatics tools are frequently utilized to analyze the results.
• Emanuele M, Costa S, Ragusa MA and Gianguzza F (*) Chromatin dynamics of the developmentally regulated P. lividus neural alpha tubulin gene. Accepted for pubblication in Int. J. Dev. Biol. (cod. nsIJDB.103264md)
• Ragusa M.A., Longo V., Emanuele M., Costa S. and Gianguzza F. (2011). In silico characterization of the neural alpha tubulin gene promoter of the sea urchin embryo Paracentrotus lividus by phylogenetic footprinting. Mol Biol Rep. in press: DOI: 10.1007/s11033-011-1016-7
• Costa S., Nicosia A., RAGUSA M.A., Cefalù A.B., Pollaccia D., Noto D., et al. (2010). A novel putative interactor for the low density lipoprotein receptor cytoplasmic domain. Molecular Medicine Reports, 3 (2), 341-345.
• Nicosia, A., Emanuele, M., Ragusa, M.A., & Costa, S. (2009). Transcriptional regulation of Tubulin α2 gene in Paracentrotus lividus. Excerpts from DBCS, 16-18.
• Casano C., Savona R., Maria A. Ragusa, Bosco L., Gianguzza F. (2008) p38 MAPK activation is required for Paracentrotus lividus skeletogenesis” Caryologia (International Journal of Cytology, Cytosystematics and Cytogenetics). 61(1): 74-81
• RAGUSA MA, Costa S, Cefalu AB, Noto D, Fayer F, Travali S, Averna MR, Gianguzza F. (2006). RT-PCR and in situ hybridization analysis of apolipoprotein H expression in rat normal tissues. Int J Mol Med. 2006 Sep;18(3):449-55. ISSN: 1107-3756
• Costa S., RAGUSA M. A., Drago G. Casano C., Alaimo G., Guida N., Gianguzza F. (2004) Sea urchin neural 2 tubulin gene: isolation and promoter analysis. Biochem. Biophys. Research Communication. 316: 446-453. ISSN: 0006-291X
• Ragusa M.A., Longo V., Emanuele M., Costa S. and Gianguzza F. (2011). In silico characterization of the neural alpha tubulin gene promoter of the sea urchin embryo Paracentrotus lividus by phylogenetic footprinting. Mol Biol Rep. in press: DOI: 10.1007/s11033-011-1016-7
• Costa S., Nicosia A., RAGUSA M.A., Cefalù A.B., Pollaccia D., Noto D., et al. (2010). A novel putative interactor for the low density lipoprotein receptor cytoplasmic domain. Molecular Medicine Reports, 3 (2), 341-345.
• Nicosia, A., Emanuele, M., Ragusa, M.A., & Costa, S. (2009). Transcriptional regulation of Tubulin α2 gene in Paracentrotus lividus. Excerpts from DBCS, 16-18.
• Casano C., Savona R., Maria A. Ragusa, Bosco L., Gianguzza F. (2008) p38 MAPK activation is required for Paracentrotus lividus skeletogenesis” Caryologia (International Journal of Cytology, Cytosystematics and Cytogenetics). 61(1): 74-81
• RAGUSA MA, Costa S, Cefalu AB, Noto D, Fayer F, Travali S, Averna MR, Gianguzza F. (2006). RT-PCR and in situ hybridization analysis of apolipoprotein H expression in rat normal tissues. Int J Mol Med. 2006 Sep;18(3):449-55. ISSN: 1107-3756
• Costa S., RAGUSA M. A., Drago G. Casano C., Alaimo G., Guida N., Gianguzza F. (2004) Sea urchin neural 2 tubulin gene: isolation and promoter analysis. Biochem. Biophys. Research Communication. 316: 446-453. ISSN: 0006-291X
No projects are available to students for the current accademic year.