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Inicio Allergologia et Immunopathologia Dissecting the localization of lipopolysaccharide-responsive and beige-like anch...
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Vol. 48. Issue 1.
Pages 8-17 (January - February 2020)
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Vol. 48. Issue 1.
Pages 8-17 (January - February 2020)
Original Article
DOI: 10.1016/j.aller.2019.07.011
Dissecting the localization of lipopolysaccharide-responsive and beige-like anchor protein (LRBA) in the endomembrane system
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Catalina Martinez-Jaramillo
Corresponding author
catalina.martinezj@udea.edu.co

Corresponding author.
, Claudia M. Trujillo-Vargas
Grupo de Inmunodeficiencias Primarias, Facultad de Medicina, Universidad de Antioquia UdeA, Calle 62 #52-59, Laboratorio 530, Sede de Investigación Universitaria, Medellín, Colombia
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Abstract
Introduction and objectives

LRBA deficiency is caused by loss of LRBA protein expression, due to either homozygous or compounds heterozygous mutations in LRBA. LRBA deficiency has been shown to affect vesicular trafficking and autophagy. To date, LRBA has been observed in the cytosol, Golgi apparatus and some lysosomes in LPS-stimulated murine macrophages. The objectives of the present study were to study the LRBA localization in organelles involved in vesicular traffic, phagocytosis, and autophagy in mononuclear phagocytes (MP).

Materials and methods

We analyzed LRBA colocalization with different endosomes markets using confocal microscopy in MP. We used the autophagy inhibitors to determine the role of LRBA in formation, maturation or degradation of the autophagosome.

Results

LRBA intracellular trafficking depends on the activity of the GTPase ADP ribosylation factor-1 (ARF) in MP. LRBA was identified in early, late endosomes but did not colocalize strongly with lysosomal markers. Although LRBA appears not to be recruited during the phagocytic cargo uptake, it greatly colocalized with the microtubule-associated protein 1A/1B-light chain 3 (LC3) under a steady state and this decreased after the induction of autophagy flux. Although the use of inhibitors of lysosome fusion did not restore the LRBA/LC3 colocalization, inhibitors of either early to late endosomes trafficking or PI3K pathway did.

Conclusions

Taken together, our results show that LRBA is located in endomembrane system vesicles, mainly in the early and late endosomes. Although LRBA appears not to be involved in the phagocytic uptake, it is recruited in the early steps of the autophagy flux.

Keywords:
LRBA
Endosomes
Phagocytosis
Autophagy
Vesicular trafficking
LC3

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