Shigella protein IpaH(9.8) is secreted from bacteria within mammalian cells and transported to the nucleus.
Various pathogenic bacteria such as Shigella deliver effector proteins into mammalian cells via the type III secretion system. The delivered Shigella effectors have been shown to variously affect host functions required for efficient bacterial internalization into the cells. In the present study, we investigated the IpaH proteins for their ability to be secreted via the type III secretion system and their fate in mammalian cells. Upon incubation in a medium containing Congo red, the bacteria secrete IpaH into the medium, but secretion of IpaH occurs later than that of IpaBCD. Immunofluorescence microscopy indicated that IpaH(9.8) is secreted from intracellular bacteria and transported into the nucleus. On microinjection of the protein, intracellular IpaH(9.8) is accumulated at one place around the nucleus and transported into the nucleus. This movement seems to be dependent on the microtubule network, since nuclear accumulation of IpaH(9.8) is inhibited in cells treated with microtubule-destabilizing agents. In nuclear import assay, IpaH(9.8) was efficiently transported into the nucleus, which was completely blocked by treatment with wheat germ agglutinin. The nuclear transport of IpaH(9.8) does not depend on host cytosolic factors but is partially dependent on ATP/GTP, suggesting that, like beta-catenin, IpaH(9.8) secreted from intracellular Shigella can be transported into the nucleus.
Toyotome
T
T
Division of Bacterial Infection, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan.
Suzuki
T
T
Kuwae
A
A
Nonaka
T
T
Fukuda
H
H
Imajoh-Ohmi
S
S
Toyofuku
T
T
Hori
M
M
Sasakawa
C
C
D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism*; D004274:DNA, Recombinant; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D021381:Protein Transport; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012760:Shigella / Q000378:metabolism*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004274:DNA, Recombinant; D011994:Recombinant Proteins; C063500:ipaH protein, Shigella flexneri