SURFACE EXCLUSION SPECIFICITY OF THE TRAT LIPOPROTEIN IS DETERMINED BY SINGLE ALTERATIONS IN A 5-AMINO-ACID REGION OF THE PROTEIN

被引:25
作者
HARRISON, JL [1 ]
TAYLOR, IM [1 ]
PLATT, K [1 ]
OCONNOR, CD [1 ]
机构
[1] UNIV SOUTHAMPTON, DEPT BIOCHEM, BASSETT CRESCENT E, SOUTHAMPTON SO9 3TU, HANTS, ENGLAND
基金
英国惠康基金;
关键词
D O I
10.1111/j.1365-2958.1992.tb01462.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The TraT protein is a highly cell-surface-exposed lipoprotein specified by F-like plasmids that confers serum resistance and blocks the conjugative transfer of plasmids to cells bearing identical or closely related plasmids, a process known as surface exclusion. The TraT protein specified by the antibiotic-resistance plasmid R6-5 was purified to apparent homogeneity. When added to mating mixtures, TraT blocked the transfer of plasmids belonging to Surface Exclusion Group IV (Sfx IV) but had no significant effect on the transfer of plasmids belonging to other groups. Additionally, the purified protein had a protective effect on bacterial cells incubated in serum, indicating that it does not have to be located on the cell surface to mediate serum resistance. To localize regions of the protein that were responsible for surface exclusion specificity, the amino acid sequence of the TraT protein specified by ColB2-K98 (Sfx II) was determined by cloning and sequencing of the corresponding gene. Comparison of the derived sequence with those of the F and R100-1 proteins indicated that surface exclusion specificity of TraT is determined by single alterations in a five-amino-acid region (residues 116-120). This was confirmed by segment swapping experiments in which the specificity of the R6-5 TraT protein (Sfx IV) was switched to that of the ColB2-K98 protein (Sfx II). Our results suggest that the region defined by residues 116-120 is located on the external face of the outer membrane and interacts specifically with the donor cell in surface exclusion.
引用
收藏
页码:2825 / 2832
页数:8
相关论文
共 39 条
  • [1] A GENETIC-ANALYSIS OF F SEX FACTOR CISTRONS NEEDED FOR SURFACE EXCLUSION IN ESCHERICHIA-COLI
    ACHTMAN, M
    MANNING, PA
    KUSECEK, B
    SCHWUCHOW, S
    WILLETTS, N
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 1980, 138 (04) : 779 - 795
  • [2] CELL-CELL INTERACTIONS IN CONJUGATING ESCHERICHIA-COLI - ROLE OF TRAT PROTEIN IN SURFACE EXCLUSION
    ACHTMAN, M
    KENNEDY, N
    SKURRAY, R
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1977, 74 (11) : 5104 - 5108
  • [3] BALDWIN TO, 1984, FOCUS, V6, P7
  • [4] MONOCLONAL-ANTIBODY DETECTION OF INCF GROUP PLASMID-ENCODED TRAT PROTEIN IN CLINICAL ISOLATES OF ESCHERICHIA-COLI
    BITTERSUERMANN, D
    PETERS, H
    JURS, M
    NEHRBASS, R
    MONTENEGRO, M
    TIMMIS, KN
    [J]. INFECTION AND IMMUNITY, 1984, 46 (02) : 308 - 313
  • [5] CHOU PY, 1979, BIOPHYS J, V26, P367, DOI 10.1016/S0006-3495(79)85259-5
  • [6] INHIBITION OF FERTILITY BY MULTIPLE DRUG-RESISTANCE FACTOR (R) IN ESCHERICHIA COLI K-12
    EGAWA, R
    HIROTA, Y
    [J]. JAPANESE JOURNAL OF GENETICS, 1962, 37 (01): : 66 - &
  • [7] INTERACTIONS BETWEEN DE-REPRESSED F-LIKE R-FACTORS AND WILD TYPE COLICIN-B FACTORS - SUPERINFECTION IMMUNITY AND REPRESSOR SUSCEPTIBILITY
    ERYDMAN, A
    MEYNELL, E
    [J]. GENETICAL RESEARCH, 1969, 14 (03) : 315 - &
  • [8] EXPERIMENTALLY IMPROVED RELIABILITY OF ULTRASENSITIVE SILVER STAINING OF PROTEIN IN POLYACRYLAMIDE GELS
    ESCHENBRUCH, M
    BURK, RR
    [J]. ANALYTICAL BIOCHEMISTRY, 1982, 125 (01) : 96 - 99
  • [9] NUCLEOTIDE-SEQUENCE OF THE SURFACE EXCLUSION GENES TRAS AND TRAT FROM THE INCF0 LAC PLASMID-PED208
    FINLAY, BB
    PARANCHYCH, W
    [J]. JOURNAL OF BACTERIOLOGY, 1986, 166 (03) : 713 - 721
  • [10] CHARACTERIZATION AND SEQUENCE-ANALYSIS OF PILIN FROM F-LIKE PLASMIDS
    FROST, LS
    FINLAY, BB
    OPGENORTH, A
    PARANCHYCH, W
    LEE, JS
    [J]. JOURNAL OF BACTERIOLOGY, 1985, 164 (03) : 1238 - 1247