Golgi polarity does not correlate with speed or persistence of freely migrating fibroblasts

Abstract

The polarization of the Golgi has long been thought to be important for cell migration. Here we show that Rat2 cells at the edge of an artificial wound repolarize the Golgi relative to the nucleus to face the direction of migration into the wound. However, in the absence of cues from neighboring cells, individual cells do not display Golgi polarity relative to the direction in which they are moving. Instead, the positioning of the Golgi relative to the nucleus remains relatively constant over time and does not reflect changes in the direction of migration. Consistent with this observation, we observe only a slight bias in Golgi positioning to the front of the nucleus and this bias is not higher during periods of time when the cell is moving in a persistent manner. Taken together, these data suggest that Golgi polarity is not a requirement for cell migration.

Nck enables directional cell migration through the coordination of polarized membrane protrusion with adhesion dynamics

“…In polarized cells migrating in two dimensions after the wounding of confluent monolayers (Uetrecht and Bear, 2009), the Golgi complex is usually positioned between the leading edge and the nucleus. As shown in Fig.…”

Cellular functions of FAK kinases: insight into molecular mechanisms and novel functions

“…There is some evidence suggesting that this event is important for directional motility into the wound; perturbation of proteins whose primary function is regulation of Golgi structure alters relocalization of the Golgi and impairs cell migration in a wound-healing assay (Yadav et al, 2009). Whether this is a universal event associated with directional motility is clearly questionable, as there is no correlation between Golgi location and direction of movement in randomly migrating fibroblasts (Uetrecht and Bear, 2009). Regardless, there is a defect in relocalization of the Golgi in wounded monolayers of cells lacking FAK (Tilghman et al, 2005;).…”

Gα12Structural Determinants of Hsp90 Interaction Are Necessary for Serum Response Element–Mediated Transcriptional Activation

“…These halfreactions were then combined and subjected to 15 additional PCR cycles. Concertina mutants in the internal ribosomal entry site (IRES) vector pLL-5.5 (Uetrecht and Bear, 2009) were engineered by first excising the Concertina cDNA from pLL-5.5 using ApaI and EcoRI, subcloning into a truncated pGEX-2T vector (GE Healthcare, Little Chalfont, Buckinghamshire, UK) at these sites and then using QuikChange II reagents to introduce codon substitutions. Mutants were subcloned into pLL-5.5 and then confirmed by sequencing.…”

“…Concertina is required for cell shape changes and migration during gastrulation (Parks and Wieschaus, 1991), but its growth signaling properties are not known. Because Concertina-specific antibodies were not available, we subcloned both wild-type and constitutively active Concertina (both Q303L and R277C variants) into the mammalian cell expression vector pLL5.5 (Uetrecht and Bear, 2009) harboring an IRES to allow translation from a bicistronic mRNA, along with green fluorescent protein (GFP). Neither wild-type nor constitutively active Concertina (Q303L or R277C) stimulated SRE-luciferase when expressed ectopically in HEK293 cells, despite the expression of GFP (Fig.…”

“…7A). Each chimera was engineered to harbor an activating Gln-to-Leu mutation within the switch II region and was subcloned into the IRES vector pLL-5.5 to allow normalization of expression through GFP immunoblotting (Uetrecht and Bear, 2009). Strikingly, the only construct that stimulated SRE signaling was chimera 4, which harbors the N-terminal 275 residues of Concertina and an additional 178 residues comprising the switch regions and C terminus of Ga 12 (Fig.…”