Methods for studying the genetics, molecular biology, physiology, and pathogenesis of the streptococci PDF
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METHODS FOR STUDYING THE GENETICS, MOLECULAR BIOLOGY, PHYSIOLOGY, AND PATHOGENESIS OF THE STREPTOCOCCI Methods for studying the genetics, molecular biology, physiology, and pathogenesis of the streptococci Edited by PAULA M. FIVES-TAYLOR and DONALD J. LEBLANC Reprinted from Methods in Cell Science, Volume 20 (1-4), 1998 SPRlNGER-SClENCE+BUSINESS MEDIA, B.Y. Library of Congress Cataloging-in-Publication Data ISBN 978-90-481-5262-9 ISBN 978-94-017-2258-2 (eBook) DOl 10.1007/978-94-017-2258-2 Printed on acid-free paper All Rights reserved © 1998 Springer Science+Business Media Dordrecht Originally published by Kluwer Academic Publishers in 1998 No part of the material protected by this copyright notice may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording or by any information storage and retrieval system, without written permission from the copyright owner. Editorial: Methods for studying the genetics, molecular biology, physiology, and pathogenesis of the streptococci Paula Fives-Taylor & Donald J. DeBlanc vii-ix Dedication xi List of contributors xiii-xvi Tn917 transponson mutagenesis and marker rescue of interrupted genes of Streptococcus mutans Dennis G. Cvitkovitch, Juan A. Gutierrez, Paula J. Crowley, Laura Wojciechowski, Jeffrey D. Hillman & Arnold S. Bleiweis 1-12 Site-specific homologous recombination mutagenesis in group B streptococci Harry H. Yim & Craig E. Rubens 13-20 Targeted mutagenesis of enterococcal genes Xiang Qin, Fang Teng, Yi Xu, Kavindra V. Singh, George M. Weinstock & Barbara E. Murray 21-33 A lactococcal pWV01-based integration toolbox for bacteria Kees Leenhouts, Gerard Venema & Jan Kok 35-50 Vectors containing streptococcal bacteriophage integrases for site-specific gene insertion W. Michael McShan, Robert E. McLaughlin, Annika Nordstrand & Joseph J. Ferretti 51-57 Streptococcal integration vectors for gene inactivation and cloning Lin Tao 59-64 Induction of transformation in streptococci by synthetic competence stimulating peptides Peter Gaustad & Donald A. Morrison 65-70 Characterization of the lactococcal conjugative element pRSOl using IS946-mediated mutagenesis David A. Mills, Trevor G. Phister, Kathleen A. Baldwin, Gary M. Dunny & Larry L. McKay 71-78 Use of electroportation in genetic analysis of enterococcal virulence Helmut Hirt, Yi Chen, Patrick M. Schlievert & Gary M. Dunny 79-84 Genetic transfer methods for Streptococcus sobrinus and other oral streptococci Donald J. LeBlanc, Yi-Ywan Chen, Nicole D. Duckley & Linda N. Lee 85-93 Isolation of enterococcal antigen-encoding genes from genomic libraries Yi Xu, Lingxia Jiang, Xiaomei Jin, Barbara E. Murray & George M. Weinstock 95-106 A simple microtiter plate screening assay for bacterial invasion or adherence Victor Nizet, Arnold L. Smith, Paul M. Sullam & Craig E. Rubens 107-111 End-probing: A non-radioactive approach to mapping transponson insertions Martin H. Lee, Aphakorn Nittayajarm & Craig E. Rubens 113-118 A method for mapping phage-inducible promoters for use in bacteriophage-triggered defense systems G. M. Djordjevic & T. R. Klaenhammer 119-126 Secretion of heterologous proteins by genetically engineered Streptococcus gordonii Teruaki Shiroza & Howard Kuramitsu 127-136 Examination of streptococcal gene expression in the mammalian environment W. Todd Grey, Joshua D. Lasker, Roy Curtiss III & Michael C. Hudson 137-142 vi Analysis of adherence-associated gene expression in Streptococcus parasangusis: A method for RNA isolation Eunice H. Froeliger & Paula Fives-Taylor 143-151 Development of an integrative, lacZ transcriptional-fusion plasmid vector for Streptococcus mutans and its use to isolate expressed genes Francesca Peruzzi, Patrick J. Piggot & Lolita Daneo-Moore 153-163 Use of proteomics and PCR to elucidate changes in protein expression in oral streptococci Robert G. Quivey Jr, Wendi L. Kuhnert & Roberta C. Faustoferri 165-179 The use of continuous flow bioreactors to explore gene expression and physiology of suspended and adherent populations of oral streptococci Robert A. Burne & Yi-Ywan M. Chen 181-190 In vitro systems for investigating group B streptococcal: host cell and extracellular matrix interactions Scott B. Winram, Glen S. Tamura & Craig E. Rubens 191-201 The rat model of endocarditis Cindy L. Munro 203-207 Lipoproteins and other cell-surface associated proteins in streptococci Roderick McNab & Howard F. Jenkinson 209-216 Growth of Streptococcal mutans in an iron-limiting medium Grace A. Spatafora & Meagan W. Moore 217-221 Identification of oral streptococci using PCR-based, reverse-capture, checkerboard hybridization Bruce J. Paster, Irena M. Bartoszyk & Floyd E. Dewhirst 223-231 Pulsed-field gel electrophoresis as an epidemiologic tool for enterococci and streptococci Jan E. Patterson & Cindy C. Kelly 233-239 Cell-based panning as a means to isolate phage display Fabs specific for a bacterial surface protein Aimee E. Stephenson, Paula Fives-Taylor & Robert J. Melamede 241-249 Subject index 251-256 Methods in Cell Science 20: vii-ix (1998), EDITORIAL Methods for studying the genetics, molecular biology, physiology, and pathogenesis of the streptococci Paula Fives-Taylorl & Donald J. LeBlanc2 1 Department of Microbiology and Molecular Genetics, University of Vermont, Burlington, Vermont, USA Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana, USA 2 This issue of Methods in Cell Science is devoted molecular techniques, the genetics and molecular entirely to methods, or new research tools, developed biology of the Gram-positive cocci lagged behind in and for studies of members of the bacterial family, that of the Enterobacteriaceae for many years. This Streptococcaceae. The methods included were seems somewhat ironic, since the origins of bacterial specifically designed for analysis of different species genetics and molecular biology can be traced to two belonging to what was once a single genus, seminal papers that describe the results of studies on Streptococcus, but which now includes three genera S. pneumoniae. The first of these papers, of course, of Streptococcaceae: the original genus, Strep represented the beginning of bacterial genetics, i.e., tococcus, in which are retained the ~-hemolytic the transformation experiements of Griffith, in 1928 species, such as S. pyogenes and S. agalactiae, the [5]. The second, and a major boost to both bacterial oral streptococci, e.g., S. mutans and S. parasanguis, genetics and molecular biology, was the identifica and S. pneumoniae; Enterococcus, formerly repre tion of DNA as Griffith's transforming principle, sented among the Lancefield Group D streptococci, by Avery, MacLeod, and McCarty, in 1944 [1]. and including such species as E. faecalis and E. However, very little work in streptococcal genetics faecium; and Lactococcus, formerly referred to as the was published until 30 years later, when Jacob and Group N, or dairy, streptococci, most notably L. Hobbs reported the first definitive demonstration of lactis. Many investigators conducting research on conjugation between streptococcal (enterococcal) the so-called 'pathogenic streptococci' seldom strains [6]. Despite the pioneering work presented interact with those who study dairy organisms, such in those three papers on members of the as the lactococci, and many of the former also fail Streptococcaceae, four years after the appearance of to recognize the pathogenic potential of the entero the latter paper, the ever widening gap between the cocci. Yet, it has become clear in recent years that streptococci and other bacterial groups, relative to procedures developed specifically for members of advances in genetics, molecular biology, and by that one genus are very often applicable to investigations time recombinant DNA technology, as well, was of all three genera. Thus, it is hoped that those pointed out in the inaugural address at the VIlth working with anyone streptococcal genus will find International Symposium on Streptococci and valuable information in articles written by investi Streptococcal Diseases, by Sir Robert Williams, gators involved with species classified in one of the President of the Symposium [8]. He noted that other two genera. although there were a large number of papers to be Certainly, many of the studies that have been presented on 'the fine details of the structure and conducted on the Streptococcaceae were initiated behaviour of the antigens of Streptococcus pyogenes, because of the diseases they cause, or to enhance but in comparison with most other areas of bacteri their utility from an industrial perspective. However, ology, rather little on genetics and antibiotic suscep the results of many of these investigations have tibility'. In fact, of the 156 presentations at that demonstrated a complexity among some members of meeting, only seven were concerned with genetic the family that would seem to warrant an interest in transfer mechanisms, mostly on the conjugative them in their own right, apart from, or in addition transfer of antibiotic resistance plasmids. Sir Robert to, any biomedical or industrial considerations. For jokingly referred to a stage play in London called 'No example, many of the streptococci and pneumococci sex please, we're British ... ', and to the fact that have as many as 30 multifunctional surface proteins. for most symposia on streptococci prior to the present Some of these prokaryotic organisms are capable one, one might give credence to the comparable of glycosylating proteins in a manner similar to assertion, 'no sex please, we're streptococci'. eukaryotes, a concept that just a few years ago would However, that symposium, with its seven papers on have been thought to be heresy. Clearly, there is genetic transfer presented in a single poorly attended much to be learned from these complex organisms. session, served as a new beginning for streptococcal Although many of the papers appearing in this genetics, a field which continues to grow to this day. issue describe highly sophisticated genetic and The enthusiastic and optimistic speakers at that VIll session met at dinner in the evening to discuss the tocols for gene transfer between bacterial strains via possibility of an international conference on strepto transformation (Gaustad and Morrison), conjugation coccal genetics, sometime in the future. Three years (Mills et aI.), electroporation (Hirt et aI.), and by later, in November of 1981, the first ASM mobilization or electroporation (LeBlanc et aI.). The International Conference on Streptococcal was held papers in section 3 describe methods for gene isola in Sarasota, FL, and was attended by 140 scientists tion (Xu et aI., and Nizet et al.), gene mapping (Lee from 14 countries, who had come to listen to 33 oral et aI.), and for the study of gene expression presentations, and read and discuss 44 posters (Djordejvic and Klaenhammer, Shiroza and devoted exclusively to streptococcal genetics [7]. Of Kuramitsu, and Grey et aI.). Section 4 concerns the many highlights of that meeting, two stand out studies on the regulation of gene expression at the in particular, the keynote address by Maclyn transcriptional (Froeliger and Fives-Taylor, and McCarty, titled 'Streptococci and the Birth of Peruzzi et aI.) and translational (Quivey et aI.) levels, Molecular Genetics' , and the session titled or by cells in an adherent versus planktonic state 'Development and Use of Recombinant DNA (Burne and Chen). The fifth section includes methods Technology'. There have now been a total of four for the study of parasite-host interactions in vitro International Streptococcal Genetics meetings since (Winram et aI.) and in vivo (Munro), or by analyses then, the latest in 1999 in Vichy, France, and atten of bacterial cell surface proteins (McNab and dance has exceeded 300 scientists from more than 20 Jenkinson). Section 6 contains papers that provide different countries. Many new genetic and molec tools for epidemiologic analysis, including species ular tools have been introduced at these meetings, specific isolation media (Spatafora and Moore), and and presented in publications stemming from them methods of species identification (Paster et aI.), and [2-4]. However, the streptococci are often difficult strain differentiation (Patterson and Kelly). Finally, to work with, and there are often minute, yet section 7 contains a single paper on the in vitro extremely critical details that are omitted from the production of antibodies (Stephenson et aI.). proceedings of scientific conferences, primarily Most of the papers in this issue deal with micro because of space constraints. In the articles appearing organisms that are pathogenic for humans. Such a in this issue of Methods in Cell Science devoted to host:parasite relationship involves a complex array the Streptococcaceae, the experimental methods have of highly specific molecular and cellular interactions. been written in such a manner as to permit someone A great deal of progress has been made in the unfamiliar with the field to repeat the experiments identification of the causes of epidemic diseases such presented without the need of additional sources. as tuberculosis and plague. However, many of the Two additional occurrences were noted in the diseases associated with organisms like the strepto inaugural address of the VIIth International cocci and enterococci, that can coexist with their host Symposium on Streptococci and Streptococcal indefinitely, and in some instances may even be Diseases. These included the resuscitation of S. considered normal members of the human flora, are mutans as a cause of dental caries, and an upsurge more intractable. Koch's postulates cannot be ful of interest in Lancefield group B streptococci and in filled for the most part. The real challenge is to group B infections. Recognition of these two species understand the mechanisms of the disease processes as the causes of important diseases has resulted in a of these bacteria that have evolved together with their large number of studies on their virulence over the human hosts, resulting in the elaboration of well past 20 years, and consequently, in the development developed and subtle strategies for coexistence. Some of a significant amount of the new methodology, properties of these bacteria, such as the ability to much of which will be applicable to other strepto usurp host functions, or to invade host cells and coccal species. In fact, strains of S. mutans, or one tissues, may facilitate peaceful coexistence with the or more other oral streptococcal species, and the host, or contribute to pathogenesis. Other properties, group B streptococci, were employed in the devel such as the ability to secrete toxins and other factors, opment of more than half of the methods to be and to cause tissue damage through stimulation of the described in the articles appearing in this issue. host inflammatory response, would seem to be overt The 27 papers appearing in this issue are grouped virulence traits. Clearly, whether coexistence or into seven sections. The first includes methods of pathogenesis prevails must depend on changes in the mutagenesis, which is accomplished by the use of regulation of many of these traits. How, then, does transposons (Cvitkovitch et aI.), suicide vectors (Yim one decipher the mechanisms and conditions associ and Rubens), or a combination of both (Qin et aI.). ated with such regulation? Perhaps the most impor The second section contains several different genetic tant area in need of study relates to the types of transfer, and/or host-vector systems. Three of the signaling involved in cell-cell communication; i.e., papers in this section describe new integration bacteria to bacteria, bacteria to host, host to bacteria, vectors (Leenhouts et aI., McShan et aI., and Tao), bacteria in mixed communities (biofilms), bacteria which can be used for multiple purposes, including in their environment. Significant insights into the mutagenesis, while the remaining four provide pro- evolution of microbial pathogenicity have been IX gained from studies of such properties at the molec References ular level, but much remains to be learned if the information is to provide a basis for the development 1. Avery OT, Macleod CM, McCarty M (1944). Studies of new therapeutic agents to prevent or interrupt on the chemical nature of the substance inducing microbial infections. Furthermore, a greater under transformation of pneumococcal types. Induction of transformation by a desoxyribonucleic fraction isolated stand of the role of the immune system in health and from pneumococcus type III. J Exp Med 79: 137-158. disease will be required. While advances have been 2. Dunny GM, Cleary PP, McKay LL (eds) (1991). made in understanding the immune response to Genetics and molecular biology of streptococci, lacto specific pathogens, what is still needed is an under cocci, and enterococci. Washington, DC: American standing of how the immune system influences all Society for Microbiology. the phases of disease progression. It is expected that 3. Ferretti JJ, Curtiss R III (eds) (1987). Streptococcal the advanced methods contained in this issue of genetics. Washington, DC: American Society for Methods in Cell Science will provide many of the Microbiology. tools required to obtain a better understanding of how 4. Ferretti JJ, Gilmore MS, Klaenhammer TR, Brown F, microorganisms interact with each other, their envi (eds) (1995). Developments in Biological Standard ronment, and their hosts. ization, Vol. 85. Genetics of Streptococci, enterococci and lactococci. New York: Karger. 5. Griffith F (1928). The significance of pneumococcal types. J Hygiene 27: 113-159. Acknowledgment 6. Jacob AE, Hobbs SJ (1974). Conjugal transfer of plasmid-borne multiple antibiotic resistance in The editors wish to express their thanks to Dr Eunice Streptococcus faecalis var. zymogenes. J Bacteriol 117: Froeliger for all her help in receiving and mailing 360-372. papers while one of the editors was on sabbatic 7. Schlessinger D (ed) (1982). Microbiology - 1982. leave. Washington, DC: American Society for Microbiology. 8. Williams REO, In: Parker MT (ed) (1979). Pathogenic Streptococci. Surrey: Reedbooks, Ltd. Methods in Cell Science 20: xi (1998). Dedication The editors have agreed to dedicate this special issue stated: 'Although the adhesiveness of bacteria for on methods for the study of the streptococci to the some mammalian cells was recognized as early as memory of the late Dr. Ronald Gibbons. The 1908 and selectivity was demonstrated by Duguid pioneering studies of Gibbons and coworkers with and Old (1980), an understanding of the role of the oral viridans group of streptococci were among microbial adhesion in the initiation of the infectious the first to establish the critical initial role of micro processes obtained a major stimulus through the bial adherence in the colonization of host mucosal studies of the attachment of oral bacteria to the surfaces. In the late sixties and early seventies very surfaces of the mouth'. While insights into the few papers were published that documented the molecular basis of streptococcal adhesion have ability of microorganisms to attach to human tissues continued with the studies performed over the past in a specific fashion. Dr. Gibbons first papers on the 25 years, much still remains to be discovered. topic appeared in the early seventies. He clearly showed that microorganisms must have specific adhesins for receptors on oral tissues and that this References specificity was responsible for their ecological niche. His work so dramatically influenced the field that the l. Gibbons RJ (1989). Bacterial adhesion to oral tissues: number of papers on bacterial adhesion increased A model for infectious diseases. J Dent Res 68: dramatically (Figure 1). In 1980, Drs. Ofek and 750-760. 2. Ofek I, Beachey EH (1980). Bacterial adherence. Adv Beachey, well-known for their work on the adhesion Intern Med 25: 503-532. of Gram-negative organisms, recognized the contri butions made by Gibbons and coworkers when they 500 450 - f" r 400 jl N 350 I' u 300 II r I 1\ t Selective m 250 ~ I:i ~ b Adhesion "" - r""' r "" ;0 ~ , e 200 !";' r 150 _!C1fJ~ if ~. '. .~: 100 I <. ". , 0 ~ - I I 50 . o ""'" 66 67 6869 7071 72737475 76777879 80 81 82 8384 8586 Year Figure 1. Papers published on bacterial adhesion. Methods in Cell Science 20: xiii-xvi (1998) List of contributors Kathleen A. Baldwin Dennis G. Cvitkovitch University of Minnesota University of Toronto Department of Food Science and Nutrition Dental Research Institute 1334 Eckles Avenue 124 Edward Street St. Paul, MN 55108, USA Toronto, Ontario, Canada M5G 1G 6 Irena M. Bartoszyk Forsyth Dental Center Lolita Daneo-Moore* Department of Molecular Genetics Temple University School of Medicine 140 Fenway Department of Microbiology and Immunology Boston, MA 02115, USA Philadelphia, PA 19140, USA Floyd E. Dewhirst Arnold S. Bleiweis* University of Florida Forsyth Dental Center Department of Molecular Genetics Department of Oral Biology Gainesville, FL 32610, USA 140 Fenway Boston, MA 02115, USA Nicole D. Buckley G. M. Djordjevic Department de Biochimie (Sciences) North Carolina State University Universite Laval, Quebec Department of Microbiology Canada Raleigh, NC 27695-7624, USA Robert A. Burne* Gary M. Dunny* University of Rochester University of Minnesota School of Medicine and Dentistry Medical School Departments of Dental Research, Microbiology and Department of Microbiology Immunology Minneapolis, MN 55455-0312, USA 601 Elmwood Avenue Rochester, NY 14642, USA Roberta C. Faustoferri University of Rochester Yi Chen School of Medicine and Dentistry University of Minnesota Department of Dental Research Medical School Box 611, 601 Elmwood Avenue Department of Microbiology Rochester, NY 14642, USA Minneapolis, MN 55455-0312, USA Joseph J. Ferretti* Yi-Ywan M. Chen University of Oklahoma Health Sciences Center University of Rochester Department of Microbiology and Immunology School of Medicine and Dentistry Oklahoma City, OK 73190, USA Department of Dental Research 601 Elmwood Avenue Paula M. Fives-Taylor* Rochester, NY 14642, USA University of Vermont Departments of Microbiology and Molecular Paula J. Crowley Genetics University of Florida Stafford Building Department of Oral Biology Burlington, VT 05405, USA Gainesville, FL 32610, USA Eunice H. Froeliger Roy Curtiss III University of Vermont Washington University Departments of Microbiology and Molecular Department of Biology Genetics St. Louis, MO 63130, USA Stafford Building Burlington, VT 05405, USA
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