Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
  • My Cart

Main menu

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About AAC
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts
  • My Cart

Search

  • Advanced search
Antimicrobial Agents and Chemotherapy
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About AAC
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
Mechanisms of Resistance

Genetic Organization of the Chromosome Region Surrounding mecA in Clinical Staphylococcal Strains: Role of IS431-Mediated mecI Deletion in Expression of Resistance in mecA-Carrying, Low-Level Methicillin- Resistant Staphylococcus haemolyticus

Yuki Katayama, Teruyo Ito, Keiichi Hiramatsu
Yuki Katayama
Department of Bacteriology, Faculty of Medicine, Juntendo University, Tokyo, Japan
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Teruyo Ito
Department of Bacteriology, Faculty of Medicine, Juntendo University, Tokyo, Japan
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Keiichi Hiramatsu
Department of Bacteriology, Faculty of Medicine, Juntendo University, Tokyo, Japan
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/AAC.45.7.1955-1963.2001
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • Fig. 1.
    • Open in new tab
    • Download powerpoint
    Fig. 1.

    Genetic organization of the mecA gene complex in staphylococci and the PCR primers for the detection of IS431L and IS1272 upstream of mecA. The structures of five classes of mec complexes found in MRSA and MRC-NS are shown. In all five classes, IS431R was present downstream of mecA. The class A meccomplex has the structuremecI-mecR1-mecA-IS431mec. In S. haemolyticus, the class A mec complex was accompanied by an upstream IS431 copy (IS431L). The class Bmec complex has the structure ψIS1272-ΔmecR1-mecA-IS431. Class C1 and C2 mec complexes have a deletion in the left side of the class A mec complex associated with an insertion of IS431. In class C1, IS431L is inserted in the same orientation as mecA, and in class C2, IS431Lis inserted in the opposite orientation relative to mecA. Class D has a deletion without any insertion sequence adjacent to the deletion point. Arrows above the genes indicate the direction of transcription. The arrowheads indicate the location and direction of primers (see Materials and Methods).

  • Fig. 2.
    • Open in new tab
    • Download powerpoint
    Fig. 2.

    Nucleotide sequencing analysis of meccomplexes and their surrounding regions. The arrows indicate the direction of transcription of genes. The region upstream of ΔmecR1 of S. caprae strain JA186 (class D) was compared with SCCmec of N315 (carrying the class Amec complex). Eight nucleotides flanking the IS431 copies are also shown to infer a possible relationship of individual IS copies. The single asterisk and the double asterisks indicate pairs of the same sequence. No conserved direct repeats indicating target duplication were found in the 8 bp flanking the IS431-mecA-IS431 structures (5).

  • Fig. 3.
    • Open in new tab
    • Download powerpoint
    Fig. 3.

    Detection of IS431L-mediated deletion in resistant mutants derived from S. haemolyticus SH621. Portions (3 μl) of the PCR mixture were applied to an agarose gel and subjected to electrophoresis. Lanes 1, λ-HindIII digest as a DNA molecular mass standard marker (the sizes were 23.1, 9.5, 6.6, 4.3, 2.3, and 2.2 kb from top to bottom); 2, SH621; 3 to 5, SH621-derived mutants with increased methicillin resistance. Mutants were selected with 1 (lane 3), 3 (lane 4), or 8 (lane 5) mg of methicillin per liter, respectively. (A) Primers iS-1 plus mA2 were used to detect the region between IS431L andmecA. (B) Primers mA4 plus iS-2 were used to detect the region between mecA and IS431R(IS431R). Note that there are varied band sizes only in lanes 3 and 4 in panel A.

  • Fig. 4.
    • Open in new tab
    • Download powerpoint
    Fig. 4.

    IS431L-mediated deletion of the class Amec complex. The structures of three subclasses of the class C1 deletion of the mec complex in the mutants are shown as follows: subclass C1a (IS431-ΔmecI-mecR1-mecA), subclass C1b (IS431-ΔmecR1 [retaining the MS domain]-mecA), and subclass C1c (IS431-ΔmecR1 [retaining only the 5′ end]-mecA). The flanking sequences of IS431L and inverted repeats (IR-L; underlined) are presented to show the deletion points in detail. The bold type indicates the names of the (mutant) strains and the sizes of the deletion. The arrows indicate the direction of transcription. A total of 15 mutants including all 10 mutants obtained by selection with 8 mg of methicillin per liter retained the class A mec complex. Others were classified as either one of the three class C subtypes. The asterisk in the mutant strain 33 shows that the position of IS431L was identical to that found in clinically isolated strain SH631.

  • Fig. 5.
    • Open in new tab
    • Download powerpoint
    Fig. 5.

    Population analysis of mutant strains SH621-37, SH621-34, and SH621-81 in comparison with the parent strains SH621 and N315. See Materials and Methods for the procedure of population analysis. Symbols: ⧫, SH621 (class A); ■, SH621-37 isolated with 3 mg of methicillin per liter (class C1); ●, SH621-34 isolated with 3 mg of methicillin per liter (class A); ▴, SH621-81 isolated with 8 mg of methicillin per liter (class A); ○, S. aureus N315 (class A).

Tables

  • Figures
  • Table 1.

    Synthetic oligonucleotides primers used for the analysis of mec complex structure

    Primer designationSequenceaLocalizationPositionsReference(s)
    mA2 5′-AACGTTGTAACCACCCCAAGA-3′ mecA 1461–144111,28
    mA4 5′-AGTGTATGATGAGCTATATGAGA-3′ mecA 1959–1981 28
    mA55′-CGCTCAGAAATTTGTTGTGC-3′ mecA 212–180 28
    mA6 5′-TATACCAAACCCGACAAC-3′ mecA 83–6628
    mA7 5′-ATCGTTGACGATAATAGCAATACA-3′ mecA 907–930 28
    mA8 5′-ATGTTTGGATTATCTTTATCATAT-3′ mecA 1863–1886 28
    iS-15′-ACATTAGATATTTGGTTGCGT-3′ IS431mec 566–5963
    iS-2 5′-TGAGGTTATTCAGATATTTCGATGT-3′IS431mec 767–743 3
    iS-35′-TCGGATGCTATCATTAAGCAT-3′ IS1272 1668–16882
    iS-4 5′-ACAATCTGTATTCTCAGGTCGT-3′ IS1272 1722–1701 2
    mI-1 5′-AATGGCGAAAAAGCACAACA-3′ mecI 1923–1942 10, 30
    mI-25′-GACTTGATTGTTTCCTCTGTT-3′ mecI 2403–228310, 30
    mcR2 5′-CGCTCAGAAATTTGTTGTGC-3′ mecR1 1954–1935 10, 30
    mcR3 5′-ATCTCCACGTTAATTCCATT-3′ mecR1 357–376 10, 30
    • ↵a The location of each primer is shown in Fig. 1.

  • Table 2.

    Distribution of various classes of mec complex among C-NS species

    mec complex classaNo. (%) of isolates of:
    S. aureus (n = 38)S. haemolyticus (n = 38)S. epidermidis(n = 34)S. sciuri (n = 9)S. caprae (n = 5)S. hominis (n = 2)S. capitis (n = 2)S. warneri (n = 1)
    A26 (68.4) 5 (13.2) 19 (55.9) 9 4 (80.0) 22 1
    B 12 (31.6) 2 (5.3) 14 (41.2) 00 0 0 0
    C1 0 1 (2.6) 0 0 00 0 0
    C2 0 30 (78.9) 1 (2.9) 0 00 0 0
    D 0 0 0 0 1 (20.0) 00 0
    • ↵a Corresponds to the meccomplex shown in Fig. 4.

  • Table 3.

    Correlation between methicillin MIC and the class ofmec complex carried by C-NS clinical strains

    SpeciesTotal no. of strains testedmeccomplex classaNo. of strains for which the MIC (mg/liter) is:
    248163264≧128
    S. epidermidis 16 A 1 4 56
    5 B 1 1 3
    1 C2 1
    S. haemolyticus 5 A 3 2
    2B 1 1
    1 C1 1
    30 C2 2 7 10 11 9
    S. sciuri 7 A2 4 1
    S. caprae 4 A 4
    1 D1
    S. hominis 2A 2
    S. capitis 2 A 2
    S. warneri 1 A 1
    • ↵a Corresponds to the meccomplex shown in Fig. 4.

  • Table 4.

    Structure of mec complex of and methicillin MIC for mutants derived from S. haemolyticus strain SH621

    StrainPBP2 productionaSelective methicillin concn (mg/liter)bClass of meccomplexcPCRdstructure of:MIC of methicillin (mg/liter)
    mecImecR1
    PBMS
    SH621 − A ++ + 2
    SH621-15 + 1 A+ + + 8
    SH621-16 + 1 A + ++ 4
    SH621-32 + 3 A + + + 512
    SH621-34 + 3 A + + + <1,024
    SH621-310 + 3 A + + + 512
    SH621-81+ 8 A + + + <1,024
    SH621-82 +8 A + + + <1,024
    SH621-83 + 8A + + + 1,024
    SH621-84 + 8 A ++ + 1,024
    SH621-85 + 8 A + + +<1,024
    SH621-86 + 8 A + + +<1,024
    SH621-87 + 8 A + + + 1,024
    SH621-88 + 8 A + + + 1,024
    SH621-89+ 8 A + + + <1,024
    SH621-810 +8 A + + + 512
    SH621-17 + 1 C1aΔ + + 16
    SH621-37 + 3 C1a Δ+ + 16
    SH621-38 + 3 C1a Δ + +16
    SH621-39 + 3 C1a Δ + + 16
    SH621-13 + 1 C1b − Δ + 32
    SH621-35 + 3 C1b − Δ + 64
    SH621-31 + 3 C1b − Δ + 32
    SH621-33 + 3 C1b − Δ + 16
    SH621-36 + 3 C1b − Δ + 16
    SH621-11 + 1 C1c − − Δ 16
    SH621-12 + 1 C1c − − Δ 16
    • ↵a Judged by the rapid slide latex agglutination assay kit (24).

    • ↵b The concentration of methicillin used for the selection of mutants.

    • ↵c Corresponds to the mec complex shown in Fig. 4.

    • ↵d + indicates that the size of the DNA fragment amplified by PCR was indistinguishable from that of SH621. Δ indicates that the amplified DNA fragment contained a deletion.

  • Table 5.

    Quantitative analysis of mecA transcripts in SH621 and its derivative strains with and without methicillin induction

    StrainsMIC of methicillin (mg/liter)Class ofmec complexAmt of mecA mRNA (amol)a
    NoninducedInducedb
    SH621 2A 1.48 2.66
    SH621-31 32 C1b 17.163.9
    SH621-810 512 A 1.79 39.8
    • ↵a Average of two experiments.

    • ↵b Induced with methicillin (8 mg/liter) for 30 min at 37°C.

PreviousNext
Back to top
Download PDF
Citation Tools
Genetic Organization of the Chromosome Region Surrounding mecA in Clinical Staphylococcal Strains: Role of IS431-Mediated mecI Deletion in Expression of Resistance in mecA-Carrying, Low-Level Methicillin- Resistant Staphylococcus haemolyticus
Yuki Katayama, Teruyo Ito, Keiichi Hiramatsu
Antimicrobial Agents and Chemotherapy Jul 2001, 45 (7) 1955-1963; DOI: 10.1128/AAC.45.7.1955-1963.2001

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this Antimicrobial Agents and Chemotherapy article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Genetic Organization of the Chromosome Region Surrounding mecA in Clinical Staphylococcal Strains: Role of IS431-Mediated mecI Deletion in Expression of Resistance in mecA-Carrying, Low-Level Methicillin- Resistant Staphylococcus haemolyticus
(Your Name) has forwarded a page to you from Antimicrobial Agents and Chemotherapy
(Your Name) thought you would be interested in this article in Antimicrobial Agents and Chemotherapy.
Share
Genetic Organization of the Chromosome Region Surrounding mecA in Clinical Staphylococcal Strains: Role of IS431-Mediated mecI Deletion in Expression of Resistance in mecA-Carrying, Low-Level Methicillin- Resistant Staphylococcus haemolyticus
Yuki Katayama, Teruyo Ito, Keiichi Hiramatsu
Antimicrobial Agents and Chemotherapy Jul 2001, 45 (7) 1955-1963; DOI: 10.1128/AAC.45.7.1955-1963.2001
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • MATERIALS AND METHODS
    • RESULTS
    • DISCUSSION
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

Related Articles

Cited By...

About

  • About AAC
  • Editor in Chief
  • Editorial Board
  • Policies
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • ASM Author Center
  • Submit a Manuscript
  • Article Types
  • Ethics
  • Contact Us

Follow #AACJournal

@ASMicrobiology

       

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

Copyright © 2019 American Society for Microbiology | Privacy Policy | Website feedback

Print ISSN: 0066-4804; Online ISSN: 1098-6596