Home LiteratureArticle Details
PMID: 11104681 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Susceptibility of mitogen-activated protein kinase kinase family members to proteolysis by anthrax lethal factor.

The Biochemical journal ·Vol. 352 Pt 3 ·2000-12-15 ·Pages 739-45

Vitale G, Bernardi L, Napolitani G, Mock M, Montecucco C

Abstract

The lethal factor (LF) produced by toxigenic strains of Bacillus anthracis is a Zn(2+)-endopeptidase that cleaves the mitogen-activated protein kinase kinases (MAPKKs) MEK1, MEK2 and MKK3. Using genetic and biochemical approaches, we have extended the study of LF proteolytic specificity to all known MAPKK family members and found that LF also cleaves MKK4, MKK6 and MKK7, but not MEK5. The peptide bonds hydrolysed by LF within all MAPKKs were identified. Cleavage invariably occurs within the N-terminal proline-rich region preceding the kinase domain, thus disrupting a sequence involved in directing specific protein-protein interactions necessary for the assembly of signalling complexes. Alignment of the sequences flanking the site of cleavage reveals the occurrence of some consensus motifs: position P2 and P1' are occupied by hydrophobic residues and at least one basic residue is present between P4 and P7. The implications of these findings for the biochemical activity and functional specificity of LF are discussed.

MeSH Terms
Amino Acid Sequence Antigens, Bacterial Bacillus anthracis/chemistry Bacterial Toxins/genetics,metabolism Catalysis HeLa Cells Humans MAP Kinase Kinase 3 MAP Kinase Kinase 4 Mitogen-Activated Protein Kinase Kinases/chemistry,genetics,metabolism Molecular Sequence Data Multigene Family Phylogeny Protein Binding Protein Processing, Post-Translational Protein Structure, Tertiary Protein-Tyrosine Kinases/chemistry,genetics,metabolism Sequence Alignment Signal Transduction Substrate Specificity Two-Hybrid System Techniques
Chemicals
Antigens, Bacterial Bacterial Toxins anthrax toxin Protein-Tyrosine Kinases MAP Kinase Kinase 3 MAP Kinase Kinase 4 MAP2K3 protein, human MAP2K4 protein, human Mitogen-Activated Protein Kinase Kinases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Vitale G
Centro CNR Biomembrane and Dipartimento di Scienze Biomediche, Università di Padova, Via Trieste 75, 35121 Padova, Italy. gvitale@makek.dstb.uniud.it
Bernardi L
Napolitani G
Mock M
Montecucco C
References (41)
41 references, click to expand
  1. The cytotoxic activity of Bacillus anthracis lethal factor is inhibited by leukotriene A4 hydrolase and metallopeptidase inhibitors.
    Biochem J. 1996 Dec 1;320 ( Pt 2):687-91 PMID: 8973585
  2. Purification and identification of a major activator for p38 from osmotically shocked cells. Activation of mitogen-activated protein kinase kinase 6 by osmotic shock, tumor necrosis factor-alpha, and H2O2.
    J Biol Chem. 1996 Oct 25;271(43):26981-8 PMID: 8900184
  3. Stress-activated protein kinases: activation, regulation and function.
    Cell Signal. 1997 Sep;9(6):403-10 PMID: 9376221
  4. Proteolytic inactivation of MAP-kinase-kinase by anthrax lethal factor.
    Science. 1998 May 1;280(5364):734-7 PMID: 9563949
  5. Lethal factor active-site mutations affect catalytic activity in vitro.
    Infect Immun. 1998 May;66(5):2374-8 PMID: 9573135
  6. Anthrax lethal factor cleaves the N-terminus of MAPKKs and induces tyrosine/threonine phosphorylation of MAPKs in cultured macrophages.
    Biochem Biophys Res Commun. 1998 Jul 30;248(3):706-11 PMID: 9703991
  7. Signaling by dual specificity kinases.
    Oncogene. 1998 Sep 17;17(11 Reviews):1447-55 PMID: 9779990
  8. JNKK1 organizes a MAP kinase module through specific and sequential interactions with upstream and downstream components mediated by its amino-terminal extension.
    Genes Dev. 1998 Nov 1;12(21):3369-81 PMID: 9808624
  9. The MKK7 gene encodes a group of c-Jun NH2-terminal kinase kinases.
    Mol Cell Biol. 1999 Feb;19(2):1569-81 PMID: 9891090
  10. Mitogen-activated protein kinase: conservation of a three-kinase module from yeast to human.
    Physiol Rev. 1999 Jan;79(1):143-80 PMID: 9922370
  11. Oligomerization of anthrax toxin protective antigen and binding of lethal factor during endocytic uptake into mammalian cells.
    Infect Immun. 1999 Apr;67(4):1853-9 PMID: 10085027
  12. Anthrax protective antigen: prepore-to-pore conversion.
    Biochemistry. 1999 Aug 10;38(32):10432-41 PMID: 10441138
  13. Anthrax.
    N Engl J Med. 1999 Sep 9;341(11):815-26 PMID: 10477781
  14. Inhibition of the mitogen-activated protein kinase kinase superfamily by a Yersinia effector.
    Science. 1999 Sep 17;285(5435):1920-3 PMID: 10489373
  15. The tranquilizing injection of Yersinia proteins: a pathogen's strategy to resist host defense.
    Biol Chem. 1999 Jul-Aug;380(7-8):795-802 PMID: 10494828
  16. Anthrax lethal factor cleaves MKK3 in macrophages and inhibits the LPS/IFNgamma-induced release of NO and TNFalpha.
    FEBS Lett. 1999 Nov 26;462(1-2):199-204 PMID: 10580119
  17. A conserved docking motif in MAP kinases common to substrates, activators and regulators.
    Nat Cell Biol. 2000 Feb;2(2):110-6 PMID: 10655591
  18. A specific activation of the mitogen-activated protein kinase kinase 1 (MEK1) is required for Golgi fragmentation during mitosis.
    J Cell Biol. 2000 Apr 17;149(2):331-9 PMID: 10769026
  19. Expression, crystallization and preliminary X-ray diffraction studies of recombinant Bacillus anthracis lethal factor.
    Acta Crystallogr D Biol Crystallogr. 2000 Nov;56(Pt 11):1449-51 PMID: 11053848
  20. On the size of the active site in proteases. I. Papain.
    Biochem Biophys Res Commun. 1967 Apr 20;27(2):157-62 PMID: 6035483
  21. Macrophages are sensitive to anthrax lethal toxin through an acid-dependent process.
    J Biol Chem. 1986 Jun 5;261(16):7123-6 PMID: 3711080
  22. Internalization and processing of Bacillus anthracis lethal toxin by toxin-sensitive and -resistant cells.
    J Biol Chem. 1989 Jul 5;264(19):11099-102 PMID: 2500434
  23. Mammalian Ras interacts directly with the serine/threonine kinase Raf.
    Cell. 1993 Jul 16;74(1):205-14 PMID: 8334704
  24. On the role of macrophages in anthrax.
    Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10198-201 PMID: 8234277
  25. Construction of Bacillus anthracis mutant strains producing a single toxin component.
    J Gen Microbiol. 1993 Oct;139(10):2459-63 PMID: 8254316
  26. Mutational analysis of residues in and around the active site of human fibroblast-type collagenase.
    J Biol Chem. 1994 Oct 21;269(42):26201-7 PMID: 7929334
  27. SNARE motif and neurotoxins.
    Nature. 1994 Dec 1;372(6505):415-6 PMID: 7984234
  28. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.
    Nucleic Acids Res. 1994 Nov 11;22(22):4673-80 PMID: 7984417
  29. Independent human MAP-kinase signal transduction pathways defined by MEK and MKK isoforms.
    Science. 1995 Feb 3;267(5198):682-5 PMID: 7839144
  30. Zinc content of the Bacillus anthracis lethal factor.
    FEMS Microbiol Lett. 1994 Dec 15;124(3):343-8 PMID: 7851740
  31. Anthrax toxin lethal factor contains a zinc metalloprotease consensus sequence which is required for lethal toxin activity.
    Mol Microbiol. 1994 Sep;13(6):1093-100 PMID: 7854123
  32. Components of a new human protein kinase signal transduction pathway.
    J Biol Chem. 1995 May 26;270(21):12665-9 PMID: 7759517
  33. Rabaptin-5 is a direct effector of the small GTPase Rab5 in endocytic membrane fusion.
    Cell. 1995 Nov 3;83(3):423-32 PMID: 8521472
  34. Improved high-level expression system for eukaryotic genes in Escherichia coli using T7 RNA polymerase and rare ArgtRNAs.
    Biotechniques. 1995 Aug;19(2):196-8, 200 PMID: 8527135
  35. MKK3- and MKK6-regulated gene expression is mediated by the p38 mitogen-activated protein kinase signal transduction pathway.
    Mol Cell Biol. 1996 Mar;16(3):1247-55 PMID: 8622669
  36. Characterization of the structure and function of a novel MAP kinase kinase (MKK6).
    J Biol Chem. 1996 Feb 9;271(6):2886-91 PMID: 8621675
  37. The vacuolar ATPase proton pump is required for the cytotoxicity of Bacillus anthracis lethal toxin.
    FEBS Lett. 1996 May 20;386(2-3):161-4 PMID: 8647272
  38. Cytoplasmic localization of mitogen-activated protein kinase kinase directed by its NH2-terminal, leucine-rich short amino acid sequence, which acts as a nuclear export signal.
    J Biol Chem. 1996 Aug 16;271(33):20024-8 PMID: 8702720
  39. Structural determinants of the specificity for synaptic vesicle-associated membrane protein/synaptobrevin of tetanus and botulinum type B and G neurotoxins.
    J Biol Chem. 1996 Aug 23;271(34):20353-8 PMID: 8702770
  40. Role of macrophage oxidative burst in the action of anthrax lethal toxin.
    Mol Med. 1994 Nov;1(1):7-18 PMID: 8790597
  41. Interaction of MAP kinase with MAP kinase kinase: its possible role in the control of nucleocytoplasmic transport of MAP kinase.
    EMBO J. 1997 Apr 15;16(8):1901-8 PMID: 9155016
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
Email
gvitale@makek.dstb.uniud.it
Published
2000-12-15
Pages
739-45
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1221512
Subset
IM
External Links
PubMed source PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2