Effects of cyclosporin A on contractile activity and cytoskeleton in chick embryo cardiomyocytes

Kołcz, Jacek; Drukała, Justyna; Jurkiewicz, Anna; Pfitzner, Roman; Garlicki, M; Czyż, Jarosław; Korohoda, W

doi:10.1139/o99-002

Cited by 9 publications

(2 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…7). Similar to the reported interaction between CyPA and neuronal Wiskott-Aldrich syndrome protein (N-WASP) (4), an interaction between CyPA and supervillin, which is a major F-actin-binding protein, would be consistent with the observed effects of CsA treatment on altered cytoskeleton and contractile functions (23,29).…”

Section: Discussionsupporting

confidence: 80%

A Conserved Tandem Cyclophilin-Binding Site in Hepatitis C Virus Nonstructural Protein 5A Regulates Alisporivir Susceptibility

Grisé

Frausto

Logan

et al. 2012

J Virol

View full text Add to dashboard Cite

Cyclophilin A (CyPA) and its peptidyl-prolyl isomerase (PPIase) activity play an essential role in hepatitis C virus (HCV) replication, and mounting evidence indicates that nonstructural protein 5A (NS5A) is the major target of CyPA. However, neither a consensus CyPA-binding motif nor specific proline substrates that regulate CyPA dependence and sensitivity to cyclophilin inhibitors (CPIs) have been defined to date. We systematically characterized all proline residues in NS5A domain II, low-complexity sequence II (LCS-II), and domain III with both biochemical binding and functional replication assays. A tandem cyclophilinbinding site spanning domain II and LCS-II was identified. The first site contains a consensus sequence motif of AØPXW (where Ø is a hydrophobic residue) that is highly conserved in the majority of the genotypes of HCV (six of seven; the remaining genotype has VØPXW). The second tandem site contains a similar motif, and the ØP sequence is again conserved in six of the seven genotypes. Consistent with the similarity of their sequences, peptides representing the two binding motifs competed for CyPA binding in a spot-binding assay and induced similar chemical shifts when bound to the active site of CyPA. The two prolines (P310 and P341 of Japanese fulminant hepatitis 1 [JFH-1]) contained in these motifs, as well as a conserved tryptophan in the spacer region, were required for CyPA binding, HCV replication, and CPI resistance. Together, these data provide a high-resolution mapping of proline residues important for CyPA binding and identify critical amino acids modulating HCV susceptibility to the clinical CPI Alisporivir. Hepatitis C virus (HCV) is a member of the plus-strand RNA virus family Flaviviridae, which includes several additional significant human pathogens, such as West Nile virus, dengue virus, and yellow fever virus. HCV infection alone affects more than 130 million people worldwide and imposes a significant toll on the global health care system, because the majority of patients are chronically infected and in need of a more effective therapy.Replication of the HCV genome occurs on intracellular membranes and requires the participation of multiple nonstructural proteins, including the nonstructural protein 3 (NS3)/NS4A protease, the NS3 helicase, membranous web-forming protein NS4B, the RNA-dependent RNA polymerase NS5B, and the replicase cofactor NS5A. In addition, the viral replication complex contains cellular proteins that are also essential for HCV replication (46). One of these cellular cofactors is human cyclophilin A (CyPA) (64), a cytosolic protein originally identified as the protein target of the immunosuppressive drug cyclosporine (CsA) (20). The targets of CsA and another immunosuppressive compound, FK506, are CyPs and FK506-binding proteins (FKBPs), respectively. The CsA-CyPA and the FK506-FKBP complexes bind to a common target, calcineurin, and block its phosphatase activity and T-cell activation (32). FK506 does not inhibit HCV in vitro (60), whereas derivatives of CsA,...

show abstract

Section: Discussionsupporting

confidence: 80%

A Conserved Tandem Cyclophilin-Binding Site in Hepatitis C Virus Nonstructural Protein 5A Regulates Alisporivir Susceptibility

Grisé

Frausto

Logan

et al. 2012

J Virol

View full text Add to dashboard Cite

show abstract

“…FKBP12 plays a role in excitation-contraction coupling in skeletal and cardiac muscle through its association with the ryanodine receptor (13)(14)(15) and inositol 1,4,5-trisphosphate (IP 3 )-sensitive Ca 2ϩ channels (16). Cyclosporin A also induces changes in the frequency and amplitude of the contractile activity of cardiomyocytes as well as in the shape and cytoskeletal organization, including the distribution of F-actin and ␣-actinin, of these cells (17). These observations suggest that FKBP12 as well as cyclophilin contribute to the dynamics of Ca 2ϩ handling in cardiomyocytes.…”

mentioning

confidence: 99%

Role of FK506-Binding Protein 12 in Development of the Chick Embryonic Heart

Okihara

Koide

Nagata

et al. 2001

Biochemical and Biophysical Research Communications

View full text Add to dashboard Cite

Knockdown endogenous CypA with siRNA in U2OS cells results in disruption of F-actin structure and alters tumor phenotype

Calhoun¹,

Lu²,

Song³

et al. 2008

Mol Cell Biochem

View full text Add to dashboard Cite

Cyclophilin A (CypA) was originally identified as a cytosolic protein possessing peptidyl-prolyl isomerase activity. CypA has been shown to play a pivotal role in the immune response, but little is known about other molecular mechanisms of CypA-mediated biologic events. In our present study, we demonstrate that knockdown CypA expression using RNAi in U2OS cells resulted in disruption of the F-actin structure, as well as decreased anchorage-independent growth, proliferation, and migration. Wild-type U2OS cells treated with cyclosporine A (CsA), a peptidyl-prolyl isomerase inhibitor, displayed the same phenotype as knockdown CypA cells, suggesting that the isomerase activity of CypA is required to maintain a normal phenotype. In vitro and in vivo binding assays revealed that CypA binds to N-WASP, which functions in the nucleation of actin via the Arp2/3 complex. Pulse-chase labeling study indicated an enhanced degradation of N-WASP in cell lacking CypA, suggesting that CypA is required for stabilizing N-WASP to form a N-WASP/Arp2/3 complex for the nucleation/initiation of F-actin polymerization.

show abstract

Effects of cyclosporin A on contractile activity and cytoskeleton in chick embryo cardiomyocytes

Cited by 9 publications

References 25 publications

A Conserved Tandem Cyclophilin-Binding Site in Hepatitis C Virus Nonstructural Protein 5A Regulates Alisporivir Susceptibility

A Conserved Tandem Cyclophilin-Binding Site in Hepatitis C Virus Nonstructural Protein 5A Regulates Alisporivir Susceptibility

Role of FK506-Binding Protein 12 in Development of the Chick Embryonic Heart

Knockdown endogenous CypA with siRNA in U2OS cells results in disruption of F-actin structure and alters tumor phenotype

Contact Info

Product

Resources

About