Suppression of villin expression by antisense RNA impairs brush border assembly in polarized epithelial intestinal cells.
We have used an antisense RNA strategy to investigate the role of the actin-associated protein, villin, in the brush-border morphogenesis of human intestinal CaCO2 cells. Stable expression of a cDNA encoding antisense villin RNA resulted in the permanent down-regulation of the endogenous villin message and dramatically affected brush-border assembly. Ultrastructural and immunolocalization studies revealed that epithelial cell polarity was largely maintained. However, in contrast to brush-border markers such as dipeptidyl-peptidase IV, the apical localization of sucrase-isomaltase was specifically impaired. Retransfection of the villin antisense-expressing cell line with a cDNA encoding a partial sense villin RNA restored both brush-border assembly and sucrase-isomaltase apical expression. The suggestion that brush-border morphogenesis may be important for the trafficking of certain proteins is discussed.
Costa de Beauregard
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MA
Institut Pasteur, Département de Biologie Moléculaire, Paris, France.
Pringault
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E
Robine
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S
Louvard
D
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D000199:Actins / Q000302:isolation & purification; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000502:physiology*; D002451:Cell Compartmentation / Q000502:physiology; D016764:Cell Polarity / Q000502:physiology*; D002999:Clone Cells; D003599:Cytoskeleton / Q000502:physiology; D015536:Down-Regulation; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D007422:Intestines / Q000166:cytology / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D008840:Microfilament Proteins / Q000235:genetics / Q000502:physiology*; D008854:Microscopy, Electron; D008871:Microvilli / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D009024:Morphogenesis / Q000187:drug effects; D016372:RNA, Antisense / Q000494:pharmacology*; D013394:Sucrase-Isomaltase Complex / Q000096:biosynthesis / Q000302:isolation & purification; D013489:Suppression, Genetic; D014162:Transfection
D000199:Actins; D002352:Carrier Proteins; D008840:Microfilament Proteins; D016372:RNA, Antisense; C026615:villin; D013394:Sucrase-Isomaltase Complex