MYO1A (Brush Border Myosin I) Dynamics in the Brush Border of LLC-PK1-CL4 Cells

The kidney epithelial cell line, LLC-PK 1-CL4 (CL4), forms a well ordered brush border (BB) on its apical surface. CL4 cells were used to examine the dynamics of MYO1A (M1A; formerly BB myosin I) within the BB using GFP-tagged MIA (GFP-M1A), MIA motor domain (GFP-MDIQ), and tail domain (GFP-Tail). G...

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Bibliographic Details
Published in:Biophysical journal 2002-04, Vol.82 (4), p.1869-1883
Main Authors: Tyska, M.J., Mooseker, M.S.
Format: Article
Language:English
Subjects:
Actins - metabolism
Animals
Biophysical Phenomena
Biophysics
Cells
Cloning, Molecular
Cytoskeleton - metabolism
Electrophoresis, Polyacrylamide Gel
Green Fluorescent Proteins
Humans
Kidney - cytology
Kidneys
Kinetics
LLC-PK1 Cells
Luminescent Proteins - metabolism
Microscopy, Confocal
Microscopy, Fluorescence
Microvilli - metabolism
Myosin Type I - chemistry
Myosin Type I - metabolism
Precipitin Tests
Protein Structure, Tertiary
Recombinant Fusion Proteins - metabolism
Subcellular Fractions
Swine
Time Factors
Transfection
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Summary:The kidney epithelial cell line, LLC-PK 1-CL4 (CL4), forms a well ordered brush border (BB) on its apical surface. CL4 cells were used to examine the dynamics of MYO1A (M1A; formerly BB myosin I) within the BB using GFP-tagged MIA (GFP-M1A), MIA motor domain (GFP-MDIQ), and tail domain (GFP-Tail). GFP- β-actin (GFP-Actin) was used to assess actin dynamics within the BB. GFP-M1A, GFP-Tail, but not GFP-MDIQ localized to the BB, indicating that the tail is sufficient for apical targeting of M1A. GFP-Actin targeted to all the actin domains of the cell including the BB. Fluorescence recovery after photobleaching analysis revealed that GFP-M1A and GFP-Tail turnover in the BB is rapid, ∼80% complete in
ISSN:0006-3495
1542-0086
DOI:10.1016/S0006-3495(02)75537-9