title: GEF1 is a ciliary Sec7 GEF of Tetrahymena thermophila.
abstract: Ciliary guanine nucleotide exchange factors (GEFs) potentially activate G proteins in intraflagellar transport (IFT) cargo release. Several classes of GEFs have been localized to cilia or basal bodies and shown to be functionally important in the prevention of ciliopathies, but ciliary Arl-type Sec 7 related GEFs have not been well characterized. Nair et al. [ 1999] identified a Paramecium ciliary Sec7 GEF, PSec7. In Tetrahymena, Gef1p (GEF1), tentatively identified by PSec7 antibody, possesses ciliary and nuclear targeting sequences and like PSec7 localizes to cilia and macronuclei. Upregulation of GEF1 RNA followed deciliation and subsequent ciliary regrowth. Corresponding to similar Psec7 domains, GEF1domains contain IQ-like motifs and putative PH domains, in addition to GBF/BIG canonical motifs. Genomic analysis identified two additional Tetrahymena GBF/BIG Sec7 family GEFs (GEF2, GEF3), which do not possess ciliary targeting sequences. GEF1 and GEF2 were HA modified to determine cellular localization. Cells transformed to produce appropriately truncated GEF1-HA showed localization to somatic and oral cilia, but not to macronuclei. Subtle defects in ciliary stability and function were detected. GEF2-HA localized near basal bodies but not to cilia. These results indicate that GEF1 is the resident Tetrahymena ciliary protein orthologous to PSec7. Cell Motil. Cytoskeleton 2009. (c) 2009 Wiley-Liss, Inc.
authors.affiliation: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York, USA. abell@aecom.yu.edu
authors.forename: Aaron J
authors.initials: AJ
authors.lastname: Bell
authors.forename: Charles
authors.initials: C
authors.lastname: Guerra
authors.forename: Vincent
authors.initials: V
authors.lastname: Phung
authors.forename: Saraswathy
authors.initials: S
authors.lastname: Nair
authors.forename: Raviraja
authors.initials: R
authors.lastname: Seetharam
authors.forename: Peter
authors.initials: P
authors.lastname: Satir
mesh_terms: D000818:Animals; D002923:Cilia / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D018503:Genome, Protozoan; D020662:Guanine Nucleotide Exchange Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D048648:Macronucleus / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D016808:Tetrahymena thermophila / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D020662:Guanine Nucleotide Exchange Factors; D015800:Protozoan Proteins