ABB  Vol.4 No.12 , December 2013
A ConA-like lectin isolated from Canavalia maritima seeds alters the expression of genes related to virulence and biofilm formation in Streptococcus mutans
ABSTRACT
Bacteria form biofilms as an adaptive mechanism in response to environmental changes. Streptococcus mutans is the biofilm-forming bacterium that is primarily associated with dental caries. The expression of important genes by bacteria in biofilms is different from that of planktonic cells. Lectins are proteins that bind specifically to carbohydrates and may have important biological activities on bacterial cells, acting as antibacterial and anti-biofilm agents. ConM (Canavalia maritima lectin) is a protein that is able to inhibit the planktonic growth and biofilm formation of S. mutans. In this context, this study aimed to evaluate the effects of ConM and concanavalin A (ConA) on the expression of genes related to virulence and biofilm formation in S. mutans. The results showed that ConM significantly reduced the expression of genes encoding enzymes related to adhesion, formation and regulation of biofilms. On the contrary, ConA did not alter the expression of the genes studied. Because the two lectins have a high degree of similarity, the differences in the actions of ConM and ConA may be explained by the small structural differences in the carbohydrate recognition domain of the lectins.

Cite this paper
Arruda Cavalcante, T. , Carneiro, V. , Neves, C. , Sousa Duarte, H. , Queiroz Martins, M. , Sousa Arruda, F. , Vasconcelos, M. , dos Santos, H. , Silva Cunha, R. , Cavada, B. and Teixeira, E. (2013) A ConA-like lectin isolated from Canavalia maritima seeds alters the expression of genes related to virulence and biofilm formation in Streptococcus mutans. Advances in Bioscience and Biotechnology, 4, 1073-7078. doi: 10.4236/abb.2013.412143.
References
[1]   McDougald, D., Rice, S.A., Barraud, N., Steinberg, P.D. and Kjelleberg, S. (2012) Should we stay or should we go: mechanisms and ecological consequences for biofilm dispersal. Nature Reviews Microbiology, 10, 39-50.
http://dx.doi.org/10.1038/nrmicro2695

[2]   Paquette, D.W., Brodala, N. and Williams, R.C. (2006) Risk factors for endosseous dental implant failure. Dental Clinics of North America, 50, 361-374.
http://dx.doi.org/10.1016/j.cden.2006.05.002

[3]   Dhir, S. (2013) Biofilm and dental implant: The microbial link. Journal of Indian Society of Periodontology, 17, 5-11. http://dx.doi.org/10.4103/0972-124X.107466

[4]   Buser, D. and Merickse-Stern, R. (1997) Long term evaluation of nonsubmerged ITI implants. Part 1: 8 year life table analysis of a prospective multicenter study with 2359 implants. Clinical Oral Implants Research, 8, 161-172. http://dx.doi.org/10.4103/0972-124X.107466

[5]   Marsh, P.D. (2005) Dental plaque: biological significance of a biofilm and community life-style. Journal of Clinical Periodontology, 32,7-15.
http://dx.doi.org/10.1034/j.1600-0501.1997.080302.x

[6]   Brooun, A., Liu, S. and Lewis, K.A. (2000) A Dose-Response study of antibiotic resistance in Pseudomonas aeruginosa biofilms. Antimicrobial Agents and Chemotherapy, 44, 640-646.
http://dx.doi.org/10.1111/j.1600-051X.2005.00790.x

[7]   Sauer, K. and Camper, A.K. (2001) Characterization of phenotypic changes in Pseudomonas putida in response to surface-associated growth. Journal of Bacteriology, 183, 6579-6589.
http://dx.doi.org/10.1128/AAC.44.3.640-646.2000

[8]   Shemesh, M., Tam, A. and Steinberg, D. (2007) Expression of biofilm-associated genes of Streptococcus mutans in response to glucose and sucrose. Journal of Medical Microbiology, 56, 1528-1535.
http://dx.doi.org/10.1128/JB.183.22.6579-6589.2001

[9]   Shemesh, M., Tam, A., Aharoni, R. and Steinberg, D. (2010) Genetic adaptation of Streptococcus mutans during biofilm formation on different types of surfaces. BMC Microbiology, 10, 51.
http://dx.doi.org/10.1099/jmm.0.47146-0

[10]   Wen, Z.T. and Burne, R.A. (2002) Functional genomics approach to identifying genes required for biofilm development by Streptococcus mutans. Applied and Environmental Microbiology, 68, 1196-1203.
http://dx.doi.org/10.1186/1471-2180-10-51

[11]   Bies, C., Lehr, C.M. and Woodley, J.F. (2004) Lectinmediated drug targeting: history and applications. Advanced Drug Delivery Reviews, 56, 425-435.
http://dx.doi.org/10.1128/AEM.69.1.722.2003

[12]   Rakhshandehroo, M., Stienstra, R., De Wit, N.J., Bragt, M.C., Haluzik, M., Mensink, R.P., Müller, M. and Kersten, S. (2012) Plasma mannose-binding lectin is stimulated by PPARα in humans. American Journal of Physiology-Endocrinology and Metabolism, 302, 595-602.
http://dx.doi.org/10.1016/j.addr.2003.10.030

[13]   Zappelli, C., Van Der Zwaan, C., Thijssen-Timmer, D.C., Mertens, K. and Meijer, A.B. (2012) Novel role for galectin-8 protein as mediator of coagulation factor V endocytosis by megakaryocytes. The Journal of Biological Chemistry, 287, 8327-8335.
http://dx.doi.org/10.1152/ajpendo.00299.2011

[14]   Hirabayashi, J. (2008) Concept, strategy and realization of lectin-based glycan profiling. Journal of Biochemistry, 144, 139-147. http://dx.doi.org/10.1074/jbc.M111.305151

[15]   Wong, J.H., Ng, T.B., Cheung, R.C., Ye, X.J., Wang, H.X., Lam, S.K., Lin, P., Chan, Y.S., Fang, E.F., Ngai, P.H., Xia, L.X., Ye, X.Y., Jiang, Y. and Liu, F. (2010) Proteins with antifungal properties and other medicinal applications from plants and mushrooms. Applied Microbiology and biotechnology, 87, 1221-1235.
http://dx.doi.org/10.1093/jb/mvn043

[16]   Teixeira, E.H., Napimoga, M.H., Carneiro, V.A., De Oliveira, T.M., Cunha, R.M., Havt, A., Martins, J.L., Pinto, V.P., Gonçalves, R.B. and Cavada, B.S. (2006) In vitro inhibition of streptococci binding to enamel acquired pellicle by plant lectins. Journal of Applied Microbiology, 101, 111-116.
http://dx.doi.org/10.1007/s00253-010-2690-4

[17]   Cavalcante, T.T., Da Rocha, B.A.M., Carneiro, V.A., Arruda, F.V.S., Do Nascimento, A.S.F., Sá, N.C., Nascimento, K.S., Cavada, B.S. and Teixeira, E.H. (2011) Effect of lectins from Diocleinae subtribe against oral Streptococci. Molecules, 16, 3530-3543.
http://dx.doi.org/10.1111/j.1365-2672.2006.02910.x

[18]   Laemmili, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685.
http://dx.doi.org/10.3390/molecules16053530

[19]   Ritz, M., Garenaux, A., Berge, M. and Federighi, M. (2009) Determination of rpoA as the most suitable internal control to study stress response in C. jejuni by RTqPCR and application to oxidative stress. Journal of Microbiological Methods, 76, 196-200.
http://dx.doi.org/10.1016/j.mimet.2008.10.014

[20]   Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular cloning: A laboratory manual. 2nd Edition, Cold Spring Harbor, New York.

[21]   Wen, Z.T., Yates, D., Ahn, S.J. and Burne, R.A. (2010) Biofilm formation and virulence expression by Streptococcus mutans are altered when grown in dual-species model. BMC Microbiology, 10, 111.
http://dx.doi.org/10.1186/1471-2180-10-111

[22]   Livak, K.J. and Schmittgen, T.D. (2001) Analysis of relative gene expression data using Real-Time Quantitative PCR and the 2–ΔΔCT Method. Methods, 25, 402-408.
http://dx.doi.org/10.1006/meth.2001.1262

[23]   Tsumori, H. and Kuramitsu, H. (1997) The role of the Streptococcus mutans glucosyltransferases in the sucrose-dependent attachment to smooth surfaces: Essential role of the GtfC enzyme. Oral Microbiology and Immunology, 12, 274-280.
http://dx.doi.org/10.1111/j.1399-302X.1997.tb00391.x

[24]   Banas, J.A. and Vickerman, M.M. (2003) Glucan-binding proteins of the oral streptococci. Critical Reviews in Oral Biology & Medicine, 14, 89-99.
http://dx.doi.org/10.1177/154411130301400203

[25]   Jakubovics, N.S., Strömberg, N., Van Dolleweerd, C.J., Kelly, C.G. and Jenkinson, H.F. (2005) Differential binding specificities of oral streptococcal antigen I/II family adhesins for human or bacterial ligands. Molecular Microbiology, 55, 1591-1605.
http://dx.doi.org/10.1111/j.1365-2958.2005.04495.x

[26]   Wen, Z.T., Baker, H.V. and Burne, R.A. (2006) Influence of BrpA on critical virulence attributes of Streptococcus mutans. Journal of Bacteriology, 188, 2983-2992.
http://dx.doi.org/10.1128/JB.188.8.2983-2992.2006

[27]   Merritt J., Kreth, J., Qi, F., Sullivan, R. and Shi, W. (2005) Non-disruptive, real-time analyses of the metabolic status and viability of Streptococcus mutans cells in response to antimicrobial treatments. Journal of Microbiology Methods, 61, 161-170.
http://dx.doi.org/10.1016/j.mimet.2004.11.012

[28]   Yang, D.Q., Liu, T.J., Zhou, X.D., He, K.F., Li, S. and Zhuang, H. (2005) Study on lactate dehydrogenase activity of Streptococcus mutans isolates derived from caries-active and caries-free individuals. Hua Xi Kou Qiang Yi Xue Za Zhi, 23, 116-118.

[29]   Hillman, J.D., Chen, A., Duncan, M. and Lee, S.W. (1994) Evidence that L-(+)-lactate dehydrogenase deficiency is lethal in Streptococcus mutans. Infection and Immunity, 62, 60-64. http://iai.asm.org/content/62/1/60

[30]   Bezerra, G.A., Oliveira, T.M., Moreno, F.B., de Souza, E.P., Rocha, B.A., Benevides, R.G., Delatorre, P., De Azevedo Jr., W.F. and Cavada, B.S. (2007) Structural analysis of Canavalia maritima and Canavalia gladiata lectins complexed with different dimannosides: New insights into the understanding of the structure biological activity relationship in legume lectins. Journal of Structural Biology, 160, 168-176.
http://dx.doi.org/10.1016/j.jsb.2007.07.012

[31]   Nóbrega, R.B., Rocha, B.M., Gadelha, C.A., SantiGadelha, T., Pires, A.F., Assreuy, A.M.S., Nascimento, K.S., Nagano, C.S., Sampaio, A.H., Cavada, B.S. and Delatorre, P. (2012) Structure of Dioclea virgata lectin: Relations between carbohydrate binding site and nitric oxide production. Biochimie, 94, 900-906.
http://dx.doi.org/10.1016/j.biochi.2011.12.009

[32]   de Vasconcelos, M.A., Cunha, C.O., Arruda, F.V.S., Carneiro, V.A., Mercante, F.M., do Nascimento Neto, L.G., de Sousa, G.S., Rocha, B.A.M., Teixeira, E.H., Cavada, B.S. and dos Santos, R.P. (2012) Lectin from Canavalia brasiliensis seeds (ConBr) is a valuable biotechnological tool to stimulate the growth of Rhizobium tropici in vitro. Molecules, 17, 5244-5254.
http://dx.doi.org/10.3390/molecules17055244

[33]   Bezerra, M.J.B., Rodrigues, N.V.F.C.,; Pires, A.F., Bezerra, G.A., Nobre, C.B., Alencar, K.L.L., Soares, P.M.G., Nascimento, K.S., Nagano, C.S., Martins, J.L., Gruber, K., Sampaio, A.H., Delatorre, P., Rocha, B.A.M., Assreuy, A.M.S. and Cavada, B.S. (2013) Crystal structure of Dioclea violacea lectin and a comparative study of vasorelaxant properties with Dioclea rostrata lectin. International Journal of Biochemistry & Cell Biology, 45, 807-815.
http://dx.doi.org/10.1016/j.biocel.2013.01.012

[34]   Cavada, B.S., Barbosa, T., Arruda, S., Grangeiro, T.B. and Barral-Netto, M. (2001) Revisiting proteus: Do minor changes in lectin structure matter in biological activity? Lessons from and potential biotechnological uses of the Diocleinae subtribe lectins. Current Protein & Peptide Science, 2, 123-135.
http://dx.doi.org/10.2174/1389203013381152

[35]   Bezerra, E.H., Rocha, B.A., Nagano, C.S., Bezerra, G.D., Moura, T.R., Bezerra, M.J., Benevides, R.G., Sampaio, A.H., Assreuy, A.M.S., Delatorre, P. and Cavada, B.S. (2011) Structural analysis of ConBr reveals molecular correlation between the carbohydrate recognition domain and nitric oxide release from endothelial cells. Biochemical and Biophysical Research Communications, 408, 566-570.
http://dx.doi.org/10.1016/j.bbrc.2011.04.061

 
 
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