Coordinate synthesis but discrete localization of homologous N-glycosylated proteins, CLP and CLB, in Naegleria pringsheimi flagellates.
The synchronous amoebae-to-flagellates differentiation of Naegleria pringsheimi has been used as a model system to study the formation of eukaryotic flagella. We cloned two novel genes, Clp, Class I on plasma membrane and Clb, Class I at basal bodies, which are transiently expressed during differentiation and characterized their respective protein products. CLP (2,087 amino acids) and CLB (1,952 amino acids) have 82.9% identity in their amino acid sequences and are heavily N-glycosylated, leading to an ~ 100 × 10(3)  increase in the relative molecular mass of the native proteins. In spite of these similarities, CLP and CLB were localized to distinct regions: CLP was present on the outer surface of the plasma membrane, whereas CLB was concentrated at a site where the basal bodies are assembled and remained associated with the basal bodies. Oryzalin, a microtubule toxin, inhibited the appearance of CLP on the plasma membrane, but had no effect on the concentration of CLB at its target site. These data suggest that N. pringsheimi uses separate mechanisms to transport CLP and CLB to the plasma membrane and to the site of basal body assembly, respectively.
Department of Systems Biology, Yonsei University, Seoul, 120-749, Korea.
In Keol
IK
Baek
Sunglan
S
Chung
Mi Ra
MR
Suh
Deog Su
DS
Hwang
Dongmin
D
Kang
Joohun
J
Lee
D015153:Blotting, Western; D002462:Cell Membrane / Q000737:chemistry; D003001:Cloning, Molecular; D020869:Gene Expression Profiling; D006031:Glycosylation; D008970:Molecular Weight; D009253:Naegleria / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015388:Organelles / Q000737:chemistry; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D015800:Protozoan Proteins