The MAL proteolipid is necessary for the overall apical delivery of membrane proteins in the polarized epithelial Madin-Darby canine kidney and fischer rat thyroid cell lines.
The MAL proteolipid has been recently demonstrated as being necessary for correct apical sorting of the transmembrane influenza virus hemagglutinin (HA) in Madin-Darby canine kidney (MDCK) cells. The fact that, in contrast to MDCK cells, Fischer rat thyroid (FRT) cells target the majority of glycosylphosphatidylinositol (GPI)-anchored proteins to the basolateral membrane provides us with the opportunity to determine the role of MAL in apical transport of membrane proteins under conditions in which the majority of GPI-anchored proteins are (MDCK cells) or are not (FRT cells) targeted to the apical surface. Using an antisense oligonucleotide-based strategy to deplete endogenous MAL, we have observed that correct transport of apical transmembrane proteins associated (HA) or not (exogenous neurotrophin receptor and endogenous dipeptidyl peptidase IV) with lipid rafts, as well as that of the bulk of endogenous apical membrane, takes place in FRT cells by a pathway that requires normal MAL levels. Even transport of placental alkaline phosphatase, a GPI-anchored protein that is targeted apically in FRT cells, was dependent on normal MAL levels. Similarly, in addition to the reported effect of MAL on HA transport, depletion of MAL in MDCK cells caused a dramatic reduction in the apical delivery of the GPI-anchored gD1-DAF protein, neurotrophin receptor, and the bulk of membrane proteins. These results suggest that MAL is necessary for the overall apical transport of membrane proteins in polarized MDCK and FRT cells.
Martín-Belmonte
F
F
Centro de Biología Molecular "Severo Ochoa," Universidad Autónoma de Madrid, Madrid, Spain.
Puertollano
R
R
Millán
J
J
Alonso
M A
MA
D000818:Animals; D001692:Biological Transport; D018958:CD55 Antigens / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D016764:Cell Polarity; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism; D004285:Dogs; D004847:Epithelial Cells / Q000378:metabolism; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D026901:Membrane Transport Proteins; D009185:Myelin Proteins / Q000235:genetics / Q000378:metabolism*; D062806:Myelin and Lymphocyte-Associated Proteolipid Proteins; D016376:Oligonucleotides, Antisense; D011510:Proteolipids / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D011916:Rats, Inbred F344; D017475:Receptors, Nerve Growth Factor / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013961:Thyroid Gland / Q000166:cytology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism
D018958:CD55 Antigens; D019267:Hemagglutinin Glycoproteins, Influenza Virus; C569026:MAL protein, human; C569028:Mal protein, rat; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D009185:Myelin Proteins; D062806:Myelin and Lymphocyte-Associated Proteolipid Proteins; D016376:Oligonucleotides, Antisense; D011510:Proteolipids; D017475:Receptors, Nerve Growth Factor; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C488986:glycoprotein D, Feline herpesvirus 1; C039447:glycoprotein D, Human herpesvirus 1; D018819:Dipeptidyl Peptidase 4