differential regulation of macropinocytosis by abi1hssh3bp1 isoforms微分调节的macropinocytosis abi1hssh3bp1亚型.pdfVIP

Differential Regulation of Macropinocytosis by Abi1/Hssh3bp1 Isoforms 1 1 1 1 2 3 Patrycja M. Dubielecka , Ping Cui , Xiaoling Xiong , Sajjad Hossain , Susanne Heck , Lyudmil Angelov , Leszek Kotula1* 1 Laboratory of Cell Signaling, New York Blood Center, New York, New York, United States of America, 2 Flow Cytometry Core, New York Blood Center, New York, New York, United States of America, 3 Confocal Microscopy Laboratory, New York Blood Center, New York, New York, United States of America Abstract Background: Macropinocytosis, which is a constitutive cellular process of fluid and macromolecule uptake, is regulated by actin cytoskeleton rearrangements near the plasma membrane. Activation of Rac1, which is proposed to act upstream of the actin polymerization regulatory Wave 2 complex, has been found to correlate with enhanced macropinocytosis. One of the components of the Wave 2 complex is Abi1. Multiple, alternatively spliced isoforms of Abi1 are expressed in mammalian cells, but the functional significance of the various isoforms is unknown. Principal Findings: Here, using flow cytometric assay analysis for Alexa Fluor 647, we demonstrate that Abi1 isoforms 2 and 3 differentially regulate macropinocytosis. LNCaP cells expressing isoform 3 had increased macropinocytic uptake that correlated with enhanced cell spreading and higher Rac1 activation in comparison to cells expressing isoform 2. Isoform 2 expressing cells had decreased macropinocytic uptake, but demonstrated greater sensitivity to Rac1 activation. Moreover, more isoform 2 was localized within the cytoplasm in comparison to isoform 3, which was more associated with the plasma membrane. Activated Rac1 was found to specifically bind to a site in exon 10 of isoform