Tetrahymena gene encodes a protein that is homologous with the liver-specific F-antigen and associated with membranes of the Golgi apparatus and transport vesicles.
The F-antigen is a prominent liver protein which has been extensively used in studies on natural and induced immunological tolerance. However, its intracellular localization and biological function have remained elusive. It has generally been assumed that the F-antigen is confined phylogenetically to vertebrates. Now we have cloned and characterized a gene from the ciliated protozoan Tetrahymena thermophila encoding a protein which clearly is homologous with the rat F-antigen. The coding region of the Tetrahymena F-antigen (TF-ag) gene specifies a 46,051 M(r) protein and is interrupted by three introns. In accordance with the predicted molecular mass of the TF-ag protein, antibodies raised against a cro-lacZ'-TF-ag fusion protein specifically recognized a 45,000 M(r) protein in Western blots of total T. thermophila protein. Immunoelectron microscopy demonstrated that the TF-ag is associated with membranes of the Golgi apparatus and transport vesicles pointing to a role of TF-ag in membrane trafficking. Transcription of the TF-ag gene, as determined by run-on analyses, was only detectable in growing cells, and following transfer to starvation condition pre-existing TF-ag mRNA was rapidly degraded. The abundance of the TF-ag protein, however, declined only moderately during prolonged periods of starvation demonstrating that extensive release of the TF-ag did not take place. In combination these results suggest that the TF-ag protein is a recycled constituent of the intracellular membrane network in T. thermophila.
Hummel
R
R
Department of Molecular Biology, University of Odense, Denmark.
Nørgaard
P
P
Andreasen
P H
PH
Neve
S
S
Skjødt
K
K
Tornehave
D
D
Kristiansen
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D017125:Genes, Protozoan / Q000235:genetics*; D006056:Golgi Apparatus / Q000737:chemistry / Q000648:ultrastructure; D007425:Intracellular Membranes / Q000737:chemistry* / Q000648:ultrastructure; D007519:Isoantigens / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid; D013217:Starvation; D016808:Tetrahymena thermophila / Q000235:genetics* / Q000648:ultrastructure; D014158:Transcription, Genetic
D007519:Isoantigens; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C030497:liver specific F antigen