The adhesion modulation protein, AmpA localizes to an endocytic compartment andinfluences substrate adhesion, actin polymerization and endocytosis invegetative Dictyostelium cells

BMC Cell Biology

Daphne D Blumberg¹  Nisha Basappa¹  Hoa N Cost¹  Jessica S Kelsey¹  Chere’ L Petty¹  Elizabeth F Noratel¹ 

[1] Department of Biological Sciences, University of Maryland, Baltimore County1000 Hilltop Circle, Baltimore, MD, 21250, USA

关键词: Dictyostelium discoideum;    Migration;    Substrate adhesion;    Endocytosis;    Actin polymerization;   

Others  :  856699
DOI  :  10.1186/1471-2121-13-29

 received in 2012-05-30, accepted in 2012-10-22,  发布年份 2012

【 摘 要 】

Background

AmpA is a secreted 24Kd protein that has pleiotropic effects on Dictyostelium development. Null mutants delay development at the mound stage with cells adhering too tightly to the substrate. Prestalk cells initially specify as prespore cells and are delayed in their migration to the mound apex. Extracellular AmpA can rescue these defects, but AmpA is also necessary in a cell autonomous manner for

a

nterior

l

ike

c

ells (ALCs) to migrate to the upper cup. The ALCs are only 10% of the developing cell population making it difficult to study the cell autonomous effect of AmpA on the migration of these cells. AmpA is also expressed in growing cells, but, while it contains a hydrophobic leader sequence that is cleaved, it is not secreted from growing cells. This makes growing cells an attractive system for studying the cell autonomous function of AmpA.

Results

In growing cells AmpA plays an environment dependent role in cell migration. Excess AmpA facilitates migration on soft, adhesive surfaces but hinders migration on less adhesive surfaces. AmpA also effects the level of actin polymerization. Knockout cells polymerize less actin while over expressing cells polymerize more actin than wild type. Overexpression of AmpA also causes an increase in endocytosis that is traced to repeated formation of multiple endocytic cups at the same site on the membrane. Immunofluorescence analysis shows that AmpA is found in the Golgi and colocalizes with calnexin and the slow endosomal recycling compartment marker, p25, in a perinuclear compartment. AmpA is found on the cell periphery and is endocytically recycled to the perinuclear compartment.

Conclusion

AmpA is processed through the secretory pathway and traffics to the cell periphery where it is endocytosed and localizes to what has been defined as a slow endosomal recycling compartment. AmpA plays a role in actin polymerization and cell substrate adhesion. Additionally AmpA influences cell migration in an environment dependent manner. Wild type cells show very little variation in migration rates under the different conditions examined here, but either loss or over expression of AmpA cause significant substrate and environment dependent changes in migration.

【 授权许可】

   
2012 Noratel et al.; licensee BioMed Central Ltd.

【 图 表 】

Fig.2.

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