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J Bacteriol, April 1998, p. 2093-2101, Vol. 180, No. 8
Department of Microbiology, University of
Alabama at Birmingham, Birmingham, Alabama
35294,1 and
Institut Biochemie,
Universität Erlangen
Received 21 November 1997/Accepted 9 February 1998
A mutant (JY2190) of Streptococcus pneumoniae Rx1 which
had acquired the ability to grow in the absence of choline and analogs was isolated. Lipoteichoic acid (LTA) and wall teichoic acid (TA) isolated from the mutant were free of phosphocholine and other phosphorylated amino alcohols. Both polymers showed an
unaltered chain structure and, in the case of LTA, an unchanged
glycolipid anchor. The cell wall composition was also not altered
except that, due to the lack of phosphocholine, the phosphate content of cell walls was half that of the parent strain. Isolated cell walls
of the mutant were resistant to hydrolysis by pneumococcal autolysin
(N-acetylmuramyl-L-alanine amidase) but were
cleaved by the muramidases CPL and cellosyl. The lack of active
autolysin in the mutant cells became apparent by impaired cell
separation at the end of cell division and by resistance against
stationary-phase and penicillin-induced lysis. As a result of the
absence of choline in the LTA, pneumococcal surface protein A (PspA)
was no longer retained on the cytoplasmic membrane. During growth in
the presence of choline, which was incorporated as phosphocholine into
LTA and TA, the mutant cells separated normally, did not release PspA, and became penicillin sensitive. However, even under these conditions, they did not lyse in the stationary phase, and they showed poor reactivity with antibody to phosphocholine and an increased release of
C-polysaccharide from the cell. In contrast to ethanolamine-grown parent cells (A. Tomasz, Proc. Natl. Acad. Sci. USA
59:86-93, 1968), the choline-free mutant cells retained
the capability to undergo genetic transformation but, compared to Rx1,
with lower frequency and at an earlier stage of growth. The properties
of the mutant could be transferred to the parent strain by DNA of the
mutant.
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Generation and Properties of a Streptococcus
pneumoniae Mutant Which Does Not Require Choline or Analogs
for Growth
Nürnberg, D-91054 Erlangen,
Germany2
*
Corresponding author. Mailing address for Werner
Fischer: Institut Biochemie, Fahrstrasse 17, D-91054 Erlangen, Germany.
Phone: 49 911-40 59 57. Fax: 49 9131-85-4605. E-mail:
w.fischer{at}biochem.uni-erlangen.de. Mailing address for
Janet Yother: BBRB 661/12, 845 19th St. S., University of
Alabama at Birmingham, Birmingham, AL 35294. Phone: (205)
934-9531. Fax: (205) 975-6715. E-mail: jyother{at}uab.edu.
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