Biochemical and cell biological characterization of a mammalian septin, Sept11.
Septins are a family of conserved cytoskeletal GTPases implicated in a variety of cellular functions such as cytokinesis and vesicle trafficking. Here, we report identification of an yet uncharacterized septin, Sept11, in septin complexes purified from porcine brain. The transcripts were detected in all tested tissues except leukocytes. A Sept11 mutant with apparently reduced GTPase activity did not form filaments in the transient expression system using COS7 cells. By Western blot analysis using a specific antibody, Sept11 was detected in various cell lines as well as brain tissues. Septin complexes immunoisolated from porcine brain with anti-Sept9 and anti-Sept11 antibodies were found to contain different Sept9 isoforms based on SDS-PAGE analyses followed by silver-staining and Western blotting. Immunofluorescent study revealed cell type-dependent intracellular localization of the protein; Sept11 was colocalized dominantly with microtubules and actin stress fibers in HMEC cells and REF52 cells, respectively, and their filamentous distribution was dependent on the cytoskeleton structures with which the protein is colocalized. Sept11 partially colocalized with stress fibers and microtubules in HeLa cells.
Hanai
Nobuhiro
N
Division of Biochemistry, Aichi Cancer Center Research Institute, 1-1 Kanokoden, Chikusa-ku, Nagoya 464-8681, Japan.
Nagata
Koh-ichi
K
Kawajiri
Aie
A
Shiromizu
Takashi
T
Saitoh
Noriko
N
Hasegawa
Yasuhisa
Y
Murakami
Shingo
S
Inagaki
Masaki
M
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001921:Brain / Q000201:enzymology; D019556:COS Cells; D020558:GTP Phosphohydrolases / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013552:Swine
D000906:Antibodies; D011994:Recombinant Proteins; D020558:GTP Phosphohydrolases