AKAP350 Is Involved in the Development of Apical "canalicular" Structures in Hepatic Cells HepG2

Overview

Journal J Cell Physiol

Specialties Cell Biology
Physiology

Date 2011 Mar 5

PMID 21374596

Citations 9

Authors

Stella M Mattaloni

Elena Kolobova

Cristian Favre

Raul A Marinelli

James R Goldenring

Maria C Larocca

Affiliations

Soon will be listed here.

Abstract

Hepatocytes are epithelial cells whose apical poles constitute the bile canaliculi. The establishment and maintenance of canalicular poles is a finely regulated process that dictates the efficiency of primary bile secretion. Protein kinase A (PKA) modulates this process at different levels. AKAP350 is an A-kinase anchoring protein that scaffolds protein complexes involved in modulating the dynamic structures of the Golgi apparatus and microtubule cytoskeleton, facilitating microtubule nucleation at this organelle. In this study, we evaluated whether AKAP350 is involved in the development of bile canaliculi-like structures in hepatocyte derived HepG2 cells. We found that AKAP350 recruits PKA to the centrosomes and Golgi apparatus in HepG2 cells. De-localization of AKAP350 from these organelles led to reduced apical cell polarization. A decrease in AKAP350 expression inhibited the formation of canalicular structures and impaired F-actin organization at canalicular poles. Furthermore, loss of AKAP350 expression led to diminished polarized expression of the p-glycoprotein (MDR1/ABCB1) at the apical "canalicular" membrane. AKAP350 knock down effects on canalicular structures formation and actin organization could be mimicked by inhibition of Golgi microtubule nucleation by depletion of CLIP associated proteins (CLASPs). Our data reveal that AKAP350 participates in mechanisms which determine the development of canalicular structures as well as accurate canalicular expression of distinct proteins and actin organization, and provide evidence on the involvement of Golgi microtubule nucleation in hepatocyte apical polarization.

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References

Kipp H, Arias I . Intracellular trafficking and regulation of canalicular ATP-binding cassette transporters. Semin Liver Dis. 2000; 20(3):339-51. DOI: 10.1055/s-2000-9388. View

Kikuchi S, Hata M, Fukumoto K, Yamane Y, Matsui T, Tamura A . Radixin deficiency causes conjugated hyperbilirubinemia with loss of Mrp2 from bile canalicular membranes. Nat Genet. 2002; 31(3):320-5. DOI: 10.1038/ng905. View

Kolobova E, Efimov A, Kaverina I, Rishi A, Schrader J, Ham A . Microtubule-dependent association of AKAP350A and CCAR1 with RNA stress granules. Exp Cell Res. 2008; 315(3):542-55. PMC: 2788823. DOI: 10.1016/j.yexcr.2008.11.011. View

Mimori-Kiyosue Y, Grigoriev I, Lansbergen G, Sasaki H, Matsui C, Severin F . CLASP1 and CLASP2 bind to EB1 and regulate microtubule plus-end dynamics at the cell cortex. J Cell Biol. 2005; 168(1):141-53. PMC: 2171674. DOI: 10.1083/jcb.200405094. View

Grindstaff K, Bacallao R, Nelson W . Apiconuclear organization of microtubules does not specify protein delivery from the trans-Golgi network to different membrane domains in polarized epithelial cells. Mol Biol Cell. 1998; 9(3):685-99. PMC: 25297. DOI: 10.1091/mbc.9.3.685. View

Shanks R, Steadman B, Schmidt P, Goldenring J . AKAP350 at the Golgi apparatus. I. Identification of a distinct Golgi apparatus targeting motif in AKAP350. J Biol Chem. 2002; 277(43):40967-72. DOI: 10.1074/jbc.M203307200. View

Muniz M, Martin M, Hidalgo J, Velasco A . Protein kinase A activity is required for the budding of constitutive transport vesicles from the trans-Golgi network. Proc Natl Acad Sci U S A. 1998; 94(26):14461-6. PMC: 25021. DOI: 10.1073/pnas.94.26.14461. View

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

Roelofsen H, Soroka C, Keppler D, Boyer J . Cyclic AMP stimulates sorting of the canalicular organic anion transporter (Mrp2/cMoat) to the apical domain in hepatocyte couplets. J Cell Sci. 1998; 111 ( Pt 8):1137-45. DOI: 10.1242/jcs.111.8.1137. View

10.

Steadman B, Schmidt P, Shanks R, Lapierre L, Goldenring J . Transforming acidic coiled-coil-containing protein 4 interacts with centrosomal AKAP350 and the mitotic spindle apparatus. J Biol Chem. 2002; 277(33):30165-76. DOI: 10.1074/jbc.M201914200. View

11.

Miller P, Folkmann A, Maia A, Efimova N, Efimov A, Kaverina I . Golgi-derived CLASP-dependent microtubules control Golgi organization and polarized trafficking in motile cells. Nat Cell Biol. 2009; 11(9):1069-80. PMC: 2748871. DOI: 10.1038/ncb1920. View

12.

Wong W, Scott J . AKAP signalling complexes: focal points in space and time. Nat Rev Mol Cell Biol. 2004; 5(12):959-70. DOI: 10.1038/nrm1527. View

13.

Rivero S, Cardenas J, Bornens M, Rios R . Microtubule nucleation at the cis-side of the Golgi apparatus requires AKAP450 and GM130. EMBO J. 2009; 28(8):1016-28. PMC: 2683699. DOI: 10.1038/emboj.2009.47. View

14.

Kumar S, ODowd C, Dunckley M, Lund T . A comparative evaluation of three transfection procedures as assessed by resistance to G418 conferred to HEPG2 cells. Biochem Mol Biol Int. 1994; 32(6):1059-66. View

15.

Larocca M, Shanks R, Tian L, Nelson D, Stewart D, Goldenring J . AKAP350 interaction with cdc42 interacting protein 4 at the Golgi apparatus. Mol Biol Cell. 2004; 15(6):2771-81. PMC: 420101. DOI: 10.1091/mbc.e03-10-0757. View

16.

Gillingham A, Munro S . The PACT domain, a conserved centrosomal targeting motif in the coiled-coil proteins AKAP450 and pericentrin. EMBO Rep. 2001; 1(6):524-9. PMC: 1083777. DOI: 10.1093/embo-reports/kvd105. View

17.

van Ijzendoorn S, Hoekstra D . Polarized sphingolipid transport from the subapical compartment changes during cell polarity development. Mol Biol Cell. 2000; 11(3):1093-101. PMC: 14833. DOI: 10.1091/mbc.11.3.1093. View

18.

Wojtal K, de Vries E, Hoekstra D, van Ijzendoorn S . Efficient trafficking of MDR1/P-glycoprotein to apical canalicular plasma membranes in HepG2 cells requires PKA-RIIalpha anchoring and glucosylceramide. Mol Biol Cell. 2006; 17(8):3638-50. PMC: 1525225. DOI: 10.1091/mbc.e06-03-0230. View

19.

Schmoranzer J, Simon S . Role of microtubules in fusion of post-Golgi vesicles to the plasma membrane. Mol Biol Cell. 2003; 14(4):1558-69. PMC: 153122. DOI: 10.1091/mbc.e02-08-0500. View

20.

Efimov A, Kharitonov A, Efimova N, Loncarek J, Miller P, Andreyeva N . Asymmetric CLASP-dependent nucleation of noncentrosomal microtubules at the trans-Golgi network. Dev Cell. 2007; 12(6):917-30. PMC: 2705290. DOI: 10.1016/j.devcel.2007.04.002. View