Cholangiocyte myosin IIB is required for localized aggregation of sodium glucose cotransporter 1 to sites of Cryptosporidium parvum cellular invasion and facilitates parasite internalization.
Internalization of the obligate intracellular apicomplexan parasite, Cryptosporidium parvum, results in the formation of a unique intramembranous yet extracytoplasmic niche on the apical surfaces of host epithelial cells, a process that depends on host cell membrane extension. We previously demonstrated that efficient C. parvum invasion of biliary epithelial cells (cholangiocytes) requires host cell actin polymerization and localized membrane translocation/insertion of Na(+)/glucose cotransporter 1 (SGLT1) and of aquaporin 1 (Aqp1), a water channel, at the attachment site. The resultant localized water influx facilitates parasite cellular invasion by promoting host-cell membrane protrusion. However, the molecular mechanisms by which C. parvum induces membrane translocation/insertion of SGLT1/Aqp1 are obscure. We report here that cultured human cholangiocytes express several nonmuscle myosins, including myosins IIA and IIB. Moreover, C. parvum infection of cultured cholangiocytes results in the localized selective aggregation of myosin IIB but not myosin IIA at the region of parasite attachment, as assessed by dual-label immunofluorescence confocal microscopy. Concordantly, treatment of cells with the myosin light chain kinase inhibitor ML-7 or the myosin II-specific inhibitor blebbistatin or selective RNA-mediated repression of myosin IIB significantly inhibits (P < 0.05) C. parvum cellular invasion (by 60 to 80%). Furthermore ML-7 and blebbistatin significantly decrease (P < 0.02) C. parvum-induced accumulation of SGLT1 at infection sites (by approximately 80%). Thus, localized actomyosin-dependent membrane translocation of transporters/channels initiated by C. parvum is essential for membrane extension and parasite internalization, a phenomenon that may also be relevant to the mechanisms of cell membrane protrusion in general.
O'Hara
Steven P
SP
Miles and Shirley Fiterman Center for Digestive Diseases, Division of Gastroenterology and Hepatology, Mayo Clinic Center for Cell Signaling, Mayo Clinic, Rochester, Minnesota 55905, USA. ohara.steven@mayo.edu
Gajdos
Gabriella B
GB
Trussoni
Christy E
CE
Splinter
Patrick L
PL
LaRusso
Nicholas F
NF
D051398:Aquaporin 1 / Q000502:physiology; D001652:Bile Ducts / Q000166:cytology / Q000469:parasitology; D015153:Blotting, Western; D002460:Cell Line; D003457:Cryptosporidiosis / Q000469:parasitology*; D016785:Cryptosporidium parvum / Q000472:pathogenicity* / Q000502:physiology; D005455:Fluorescent Antibody Technique; D006576:Heterocyclic Compounds, 4 or More Rings / Q000494:pharmacology; D006790:Host-Parasite Interactions / Q000502:physiology; D006801:Humans; D008855:Microscopy, Electron, Scanning; D024507:Nonmuscle Myosin Type IIA / Q000502:physiology; D024501:Nonmuscle Myosin Type IIB / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051269:Sodium-Glucose Transporter 1 / Q000037:antagonists & inhibitors / Q000502:physiology*
D006576:Heterocyclic Compounds, 4 or More Rings; D051269:Sodium-Glucose Transporter 1; D051398:Aquaporin 1; C472645:blebbistatin; D024507:Nonmuscle Myosin Type IIA; D024501:Nonmuscle Myosin Type IIB
Eur J Cell Biol. 1999 Nov;78(11):794-801
10604656
Microbes Infect. 2002 Aug;4(10):1047-58
12191655
J Am Soc Nephrol. 2006 Jan;17(1):39-45
16319186
J Biol Chem. 1997 May 16;272(20):12984-8
9148905
Biochem Biophys Res Commun. 2005 Jun 17;331(4):1560-5
15883051
Bioessays. 1996 Mar;18(3):179-82
8867731
Mol Biol Cell. 2004 Feb;15(2):688-95
14617807
Cell Microbiol. 2006 Dec;8(12):1932-45
16848787
J Biol Chem. 2006 Nov 24;281(47):35873-83
17020881
J Biol Chem. 2009 Dec 18;284(51):35273-7
19846560
Parasitol Res. 1988;74(6):531-6
3194366
J Parasitol. 2004 Apr;90(2):212-21
15165040
J Protozool. 1991 Nov-Dec;38(6):82S-84S
1818219
Parasitol Res. 2004 Mar;92(4):317-27
14727189
Biochem J. 2006 Dec 1;400(2):225-34
16848764
J Cell Biol. 1996 Jun;133(6):1403-15
8682874
J Parasitol. 2005 Oct;91(5):995-9
16419739
J Biol Chem. 2006 Aug 25;281(34):24662-70
16790446
J Biol Chem. 2007 Jul 27;282(30):22102-11
17519229
Nature. 2005 Apr 7;434(7034):786-92
15815633
Curr Opin Cell Biol. 2004 Feb;16(1):61-7
15037306
Semin Liver Dis. 2002 Aug;22(3):277-89
12360421
J Biol Chem. 2005 May 20;280(20):19594-9
15774463
Gastrointest Endosc Clin N Am. 1998 Oct;8(4):963
9730942
Infect Immun. 2005 Sep;73(9):5379-87
16113253
J Biol Chem. 2004 Jan 23;279(4):2800-8
14594953
Gastroenterology. 2000 Feb;118(2):368-79
10648465
Am J Physiol. 1994 Jun;266(6 Pt 1):G1060-70
8023938
Infect Immun. 1999 Feb;67(2):844-52
9916099
Gastroenterology. 1986 Mar;90(3):583-94
3943690
Proc Natl Acad Sci U S A. 2005 May 3;102(18):6338-43
15851691
J Cell Sci. 2005 Dec 15;118(Pt 24):5691-8
16303850
J Parasitol. 1998 Oct;84(5):908-13
9794629
J Biol Chem. 2004 Jun 25;279(26):27450-7
15069078
J Physiol. 2002 Jul 1;542(Pt 1):3-16
12096044
Cytoskeleton (Hoboken). 2010 Jan;67(1):32-42
19743471
Infect Immun. 2006 Jan;74(1):99-107
16368962
N Engl J Med. 2002 May 30;346(22):1723-31
12037153
J Biol Chem. 2003 Oct 3;278(40):38132-40
12847096
Infect Immun. 2004 May;72(5):3011-21
15102814
Gastroenterology. 2003 Jul;125(1):216-28
12851885
Curr Opin Cell Biol. 1999 Feb;11(1):26-33
10047526
J Leukoc Biol. 2002 Feb;71(2):212-22
11818441
J Biochem. 1999 Feb;125(2):354-62
9990134
Nat Cell Biol. 2005 Mar;7(3):255-61
15723050
J Biochem. 1995 Dec;118(6):1104-7
8720121
Physiol Rev. 1998 Jan;78(1):247-306
9457175
J Biol Chem. 2005 Jun 24;280(25):23741-7
15826951
Infect Immun. 2000 Apr;68(4):2315-22
10722635
J Protozool. 1986 Feb;33(1):98-108
3959014
J Parasitol. 2001 Apr;87(2):429-32
11318578
J Muscle Res Cell Motil. 1995 Aug;16(4):379-89
7499478
Biophys J. 2006 Nov 15;91(10):3907-20
16920834
Infect Immun. 2001 Sep;69(9):5940-2
11500478
Biochim Biophys Acta. 2006 Aug;1758(8):1085-93
16564496
PLoS One. 2007 Aug 01;2(7):e658
17668046
Adv Parasitol. 1998;40:5-36
9554069