JBC Ideal method for primary cell transfection

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M010287200 on March 22, 2001

J. Biol. Chem., Vol. 276, Issue 24, 21053-21061, June 15, 2001
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
276/24/21053    most recent
M010287200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Llull, D.
Right arrow Articles by López, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Llull, D.
Right arrow Articles by López, R.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Tts, a Processive beta -Glucosyltransferase of Streptococcus pneumoniae, Directs the Synthesis of the Branched Type 37 Capsular Polysaccharide in Pneumococcus and Other Gram-positive Species*

Daniel LlullDagger , Ernesto García§, and Rubens López

From the Centro de Investigaciones Biológicas, CSIC, Velázquez 144, 28006 Madrid, Spain

The type 37 capsule of Streptococcus pneumoniae is a homopolysaccharide built up from repeating units of [beta -D-Glcp-(1right-arrow2)]-beta -D-Glcp-(1right-arrow3). The elements governing the expression of the tts gene, coding for the glucosyltransferase involved in the synthesis of the type 37 pneumococcal capsular polysaccharide, have been studied. Primer extension analysis and functional tests demonstrated the presence of four new transcriptional start points upstream of the previously reported tts promoter (ttsp). Most interesting, three of these transcriptional start points are located in a RUP element thought to be involved in recombinational events (Oggioni, M. R., and Claverys, J. P. (1999) Microbiology 145, 2647-2653). Transformation experiments using either a recombinant plasmid containing the whole transcriptional unit of tts or chromosomal DNA from a type 37 pneumococcus showed that tts is the only gene required to drive the biosynthesis of a type 37 capsule in S. pneumoniae and other Gram-positive bacteria, namely Streptococcus oralis, Streptococcus gordonii, and Bacillus subtilis. The Tts synthase was overproduced in S. pneumoniae and purified as a membrane-associated enzyme. These membrane preparations used UDP-Glc as substrate to catalyze the synthesis of a high molecular weight polysaccharide immunologically identical to the type 37 capsule. In addition, UDP-Gal was also a substrate to produce type 37 polysaccharide since a strong UDP-Glc-4'-epimerase activity is associated to the membrane fraction of S. pneumoniae. These results indicated that Tts has a dual biochemical activity that leads to the synthesis of the branched type 37 polysaccharide.

* This work was supported in part by Grants PB96-0809 and BMC2000-1002 from the Dirección General de Investigación Científica y Técnica.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Recipient of a doctoral fellowship from Programa Sectorial de Formación de Profesorado Universitario y Personal Investigador (Ministerio de Educación y Cultura).

§ To whom correspondence should be addressed. Tel.: 34-91-561-1800; Fax: 34-91-562-7518; E-mail: e.garcia@cib.csic.es.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 Appl. Environ. Microbiol.Home page
M. Dols-Lafargue, H. Y. Lee, C. Le Marrec, A. Heyraud, G. Chambat, and A. Lonvaud-Funel
Characterization of gtf, a Glucosyltransferase Gene in the Genomes of Pediococcus parvulus and Oenococcus oeni, Two Bacterial Species Commonly Found in Wine
Appl. Envir. Microbiol., July 1, 2008; 74(13): 4079 - 4090.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
D. M. Aanensen, A. Mavroidi, S. D. Bentley, P. R. Reeves, and B. G. Spratt
Predicted Functions and Linkage Specificities of the Products of the Streptococcus pneumoniae Capsular Biosynthetic Loci
J. Bacteriol., November 1, 2007; 189(21): 7856 - 7876.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
R. T. Cartee, W. T. Forsee, and J. Yother
Initiation and Synthesis of the Streptococcus pneumoniae Type 3 Capsule on a Phosphatidylglycerol Membrane Anchor
J. Bacteriol., July 1, 2005; 187(13): 4470 - 4479.
[Abstract] [Full Text] [PDF]

Home page
 MicrobiologyHome page
D. Llull, P. Veiga, J. Tremblay, and S. Kulakauskas
Immobilization-based isolation of capsule-negative mutants of Streptococcus pneumoniae
Microbiology, June 1, 2005; 151(6): 1911 - 1917.
[Abstract] [Full Text] [PDF]

Home page
 J Med MicrobiolHome page
F. Kong, W. Wang, J. Tao, L. Wang, Q. Wang, A. Sabananthan, and G. L Gilbert
A molecular-capsular-type prediction system for 90 Streptococcus pneumoniae serotypes using partial cpsA-cpsB sequencing and wzy- or wzx-specific PCR
J. Med. Microbiol., April 1, 2005; 54(4): 351 - 356.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
J. K. Morona, R. Morona, D. C. Miller, and J. C. Paton
Mutational Analysis of the Carboxy-Terminal (YGX)4 Repeat Domain of CpsD, an Autophosphorylating Tyrosine Kinase Required for Capsule Biosynthesis in Streptococcus pneumoniae
J. Bacteriol., May 15, 2003; 185(10): 3009 - 3019.
[Abstract] [Full Text] [PDF]

Home page
 MicrobiologyHome page
R. D. Waite, D. W. Penfold, J. K. Struthers, and C. G. Dowson
Spontaneous sequence duplications within capsule genes cap8E and tts control phase variation in Streptococcus pneumoniae serotypes 8 and 37
Microbiology, February 1, 2003; 149(2): 497 - 504.
[Abstract] [Full Text] [PDF]

Home page
 Infect. Immun.Home page
P. Giammarinaro and J. C. Paton
Role of RegM, a Homologue of the Catabolite Repressor Protein CcpA, in the Virulence of Streptococcus pneumoniae
Infect. Immun., October 1, 2002; 70(10): 5454 - 5461.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2001 by the American Society for Biochemistry and Molecular Biology.