title: Identification of a novel nuclear localization signal sequence in Chlamydia trachomatis-secreted hypothetical protein CT311.
abstract: We previously reported that Chlamydia trachomatis hypothetical protein CT311 was secreted out of chlamydial inclusion and into host cell cytosol. We now found that CT311 further entered host cell nucleus at the late stage of infection and continued to accumulate in the nucleus of C. trachomatis-infected cells. When CT311 was expressed via a transgene in mammalian cells, CT311 protein was exclusively detected in the nucleus, suggesting that CT311 by itself is sufficient for nuclear targeting. However, preexisting nuclear CT311 did not affect subsequent chlamydial infection. Using deletion constructs, we mapped a nuclear localization signal sequence of CT311 to residues 21 to 63 ((21)AVEGKPLSRAAQLRERRKDLHVSGKPSPRYALKKRALEAKKNK(63)). This sequence was sufficient for targeting a heterologous protein into mammalian cell nucleus and it contains two independent clusters of basic residues ((34)RERRK(38) and (53)KKRALEAKKNK(63) respectively). Deletion or alanine substitution of the basic residues in either cluster led to loss of nuclear targeting activity, suggesting that both clusters are critical for the nuclear targeting function. These observations have demonstrated that the hypothetical protein CT311 possesses a novel nuclear localization signal sequence with dual modules of basic residues for targeting host cell nucleus during Chlamydia trachomatis infection.
authors.affiliation: Department of Microbiology and Immunology, University of Texas Health Science Center at San Antonio, San Antonio, Texas, United States of America.
authors.forename: Lei
authors.initials: L
authors.lastname: Lei
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Dong
authors.forename: Zhongyu
authors.initials: Z
authors.lastname: Li
authors.forename: Guangming
authors.initials: G
authors.lastname: Zhong
mesh_terms: D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D002692:Chlamydia trachomatis / Q000378:metabolism*; D006367:HeLa Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals; D010449:Peptide Mapping
chemical_list: D001426:Bacterial Proteins; D019913:Nuclear Localization Signals