A series of step-level penicillin-resistant derivatives of Streptococcus sanguis V288 (Challis) were obtained through successive genetic transformations. The DNA donor used was a laboratory-derived, penicillin-resistant multistep mutant of the recipient strain. Detection of the penicillin-binding proteins (PBPs) of wild-type and transformants revealed five major PBPs. While it was found that S. sanguis can acquire intrinsic resistance in a stepwise manner and the mechanism was similar to those of some other organisms (changes in penicillin-binding protein affinity and/or in extent of penicillin binding), multiple-PBP changes accompanied a single step-level of resistance. All of the PBPs showed varying degrees of decreased affinity for [3H]benzylpenicillin with increasing penicillin resistance. Of these, the consistent, dramatic and progressive decrease of PBP 4 binding was most notable. After an initial decrease at the first step-level of resistance, PBP 5 was restored to wild-type levels, indicating a possible important role in survival. Genetic linkage of the first two step-levels of resistance was demonstrated by examination of transformation frequencies and by hit-kinetics experiments. A convenient method is described for the quantitative comparison of fluorographs containing PBPs with a wide range of affinities for penicillin.
BlumbergP. M.,
StromingerJ. L.1974; Interaction of penicillin with the bacterial cell: penicillin-binding proteins and penicillin-sensitive enzymes. Bacteriological Reviews 38:291–335
BuchananC. E.,
StromingerJ. L.1976; Altered penicillin-binding components in penicillin-resistant mutants of Bacillus subtilis. Proceedings of the National Academy of Sciences of the United States of America 73:1816–1820
ButlerL. O.,
SmileyM. B.1970; Characterization by transformation of an ampicillin-resistant mutant of pneumococcus. Journal of General Microbiology 61:189–195
Daneo-MooreL.,
PucciM.,
ZitoE. T.,
FerreroM.1983; Studies of a beta-lactam resistance in Streptococcus faecium. In The Target of Penicillin pp. 493–498HakenbeckR.,
HöltjeJ.-V.,
LabischinskiH.
Edited by Berlin: Walter de Gruyter;
DoughertyT. J.1984; Intrinsic resistance: penicillin target alterations and effects on cell wall synthesis. In Microbiology1984 pp. 398–401LeiveL.,
SchlessingerD.
Edited by Washington, DC: American Society for Microbiology;
DoughertyT. J.,
RollerA. E.,
TomaszA.1980; Penicillin-binding proteins of penicillin- susceptible and intrinsically resistant Neisseria gonorrhoeae. Antimicrobial Agents and Chemotherapy 18:730–737
FontanaR.,
CanepariP.,
SattaG.1983; The role of a protein that binds penicillin with slow kinetics in physiology response to penicillin of Streptococcus faecium ATCC 9790. In The Target of Penicillin pp. 531–536HakenbeckR.,
HöltjeJ.-V.,
LabischinskiH.
Edited by Berlin: Walter de Gruyter;
GodfreyA. J.,
BryanL. E.1982; Mutation of Pseudomonas aeruginosa specifying reduced affinity for penicillin G. Antimicrobial Agents and Chemotherapy 21:216–223
GoodgalS. H.1961; Studies on transformation of Haemophilus influenzae. IV. Linked and unlinked transformations. Journal of General Physiology 45:205–228
HakenbeckR.,
TarpayM.,
TomaszA.1980; Multiple changes of penicillin-binding proteins in penicillin-resistant clinical isolates of Streptococcus pneumoniae. Antimicrobial Agents and Chemotherapy 17:364–371
HakenbeckR.,
EllerbrokH.,
BrieseT.,
HandwergerS.,
TomaszA.1986; Penicillin-binding proteins of penicillin-susceptible and -resistant pneumococci: immunological relatedness of altered proteins and changes in peptides carrying the β-lactam site. Antimicrobial Agents and Chemotherapy 30:553–558
HandwergerS.,
TomaszA.1986; Alterations in kinetic properties of penicillin-binding proteins of penicillin-resistant Streptococcus pneumoniae. Antimicrobial Agents and Chemotherapy 30:57–63
HartmanB. J.,
TomaszA.1984; Low-affinity penicillin-binding protein associated with β-lactam resistance in Staphylococcus aureus. Journal of Bacteriology 158:513–516
LaskeyR. A.,
MillsA. D.1975; Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. European Journal of Biochemistry 56:335–341
ReiderJ. L.,
MacrinaF. L.1976; Plasmid DNA isolation in Streptococcus mutans: glycine-enhanced lysis. In Proceedings: Microbial Aspects of Dental Caries (A Special Supplement to Microbiology Abstracts)3 pp. 725–736StilesH. M.,
LoescheW. J.,
O’BrienT. C.
Edited by Washington, DC: Information Retrieval;
RossiL.,
ToninE.,
ChengY. R.,
FontanaR.1985; Regulation of penicillin-binding protein activity: description of a methicillin-inducible penicillin-binding protein in Staphylococcus aureus. Antimicrobial Agents and Chemotherapy 27:828–831
TomaszA.,
Zighelboim-DaumS.,
HandwergerS.,
LivH.,
QianH.1984; Physiology and genetics of intrinsic beta-lactam resistance in pneumococci. In Microbiology1984 pp. 393–397LeiveL.,
SchlessingerD.
Edited by Washington, DC: American Society for Microbiology;
UbukataK.,
YamashitaN.,
KonnoM.1985; Occurrence of a β-lactam-inducible penicillin-binding protein in methicillin-resistant staphylococci. Antimicrobial Agents and Chemotherapy 30:57–63
WashingtonJ. A.IISutterV. L.1980; Dilution susceptibility test: agar and macro-broth dilution procedures. In Manual of Clinical Microbiology, 3rd edn. pp. 453–458LennetteE. H.,
BalowsA.,
HauslerW. J.JrTruantJ. P.
Edited by Washington, DC: American Society for Microbiology;
WaxmanD. J.,
StromingerJ. L.1983; Penicillinbinding proteins and the mechanisms of action of β-lactam antibiotics. Annual Review of Biochemistry 52:825–869
ZighelboimS.,
TomaszA.1980; Penicillinbinding proteins of multiply antibiotic-resistant South African strains of Streptococcus pneumoniae. Antimicrobial Agents and Chemotherapy 17:434–442
ZimmermanW.1980; Penetration of β-lactam antibiotics into their target enzymes in Pseudomonas aeruginosa: comparison of a highly sensitive mutant with its parent strain. Antimicrobial Agents and Chemotherapy 18:94–100