PMID- 15499383
OWN - NLM
STAT- MEDLINE
DCOM- 20050322
LR  - 20061115
IS  - 0829-8211 (Print)
IS  - 0829-8211 (Linking)
VI  - 82
IP  - 5
DP  - 2004 Oct
TI  - Dynamin- and clathrin-dependent endocytic pathway in unicellular eukaryote 
      Paramecium.
PG  - 547-58
AB  - The first evidence of dynamin presence and its colocalization with clathrin in 
      the compartment involved in Paramecium receptor-mediated endocytosis is 
      presented. We identified dynamin by cloning, Western blotting, and 
      immunodetection in confocal and electron microscopy. The partial genes, which we 
      have designated ParDyn1 and ParDyn2, are 1091 bp long, 90% identical to one 
      another and encode the N-terminal and middle domains of Paramecium dynamin 
      isoform 1 and isoform 2. The deduced amino acid sequences contain all three 
      guanosine 5'-triphosphate (GTP)-binding motifs and show 67% homology to mammalian 
      dynamins. Antibodies generated against the cloned GTPase domain revealed dynamin 
      association with endosomes containing transferrin, the marker of 
      receptor-mediated endocytosis. In Western blotting a strong immunoreactive 
      polypeptide of approximately 116 kDa, which seems to be phosphorylated, was 
      accompanied by a faint one of approximately 90 kDa in cytosolic fraction (S2). 
      Dynamin level was correlated with internalization of transferrin and it was 
      significantly decreased upon inhibition of this process. Immunogold labeling in 
      electron microscopy revealed colocalization of dynamin and clathrin in coated 
      pits and endocytic vesicles. Moreover, the polypeptide cross-reaction with 2 
      different antibodies against mammalian clathrin was identified by immunoblotting. 
      These results indicate that dynamin- and clathrin-dependent pathway exists in 
      this evolutionary ancient cell.
FAU - Wiejak, Jolanta
AU  - Wiejak J
AD  - Laboratory of Cell Membrane Physiology, Department of Cell Biology, Nencki 
      Institute of Experimental Biology, 3 Pasteur Str., 02-093 Warsaw, Poland.
FAU - Surmacz, Liliana
AU  - Surmacz L
FAU - Wyroba, Elzbieta
AU  - Wyroba E
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Canada
TA  - Biochem Cell Biol
JT  - Biochemistry and cell biology = Biochimie et biologie cellulaire
JID - 8606068
RN  - 0 (Clathrin)
RN  - 0 (Protozoan Proteins)
RN  - 0 (Receptors, Transferrin)
RN  - EC 3.6.5.5 (Dynamins)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Cell Membrane/metabolism
MH  - Cells, Cultured
MH  - Clathrin/*metabolism
MH  - Cloning, Molecular
MH  - Coated Pits, Cell-Membrane/metabolism
MH  - Dynamins/genetics/*metabolism
MH  - Endocytosis/*physiology
MH  - Endosomes/*metabolism
MH  - Microscopy, Immunoelectron
MH  - Molecular Sequence Data
MH  - Paramecium/*metabolism
MH  - Protozoan Proteins/genetics/metabolism
MH  - Receptors, Transferrin/metabolism
MH  - Sequence Homology, Amino Acid
EDAT- 2004/10/23 09:00
MHDA- 2005/03/23 09:00
CRDT- 2004/10/23 09:00
PHST- 2004/10/23 09:00 [pubmed]
PHST- 2005/03/23 09:00 [medline]
PHST- 2004/10/23 09:00 [entrez]
AID - o04-098 [pii]
AID - 10.1139/o04-098 [doi]
PST - ppublish
SO  - Biochem Cell Biol. 2004 Oct;82(5):547-58. doi: 10.1139/o04-098.