Retargeting from the CR3 to the LFA-1 receptor uncovers the adenylyl cyclase enzyme?translocating segment ofBordetellaadenylate cyclase toxin

0533024 - MBÚ 2021 RIV US eng J - Journal Article
Mašín, Jiří - Osičková, Adriana - Jurnečka, David - Klímová, Nela - Khaliq, Humaira - Šebo, Peter - Osička, Radim
Retargeting from the CR3 to the LFA-1 receptor uncovers the adenylyl cyclase enzyme?translocating segment ofBordetellaadenylate cyclase toxin.
Journal of Biological Chemistry. Roč. 295, č. 28 (2020), s. 9349-9365. ISSN 0021-9258. E-ISSN 1083-351X
R&D Projects: GA ČR(CZ) GA19-04607S; GA ČR(CZ) GA18-18079S; GA ČR(CZ) GA19-12695S; GA ČR(CZ) GA18-20621S; GA MŠMT(CZ) LM2018133
Institutional support: RVO:61388971
Keywords : AC domain translocation * acylation * acyltransferase * fatty acyl * integrin * RTX toxin
OECD category: Microbiology
Impact factor: 5.157, year: 2020
Method of publishing: Open access with time embargo
https://www.jbc.org/content/early/2020/05/11/jbc.RA120.013630.short

TheBordetellaadenylate cyclase toxin-hemolysin (CyaA) and the ?-hemolysin (HlyA) ofEscherichia colibelong to the family of cytolytic pore-forming Repeats in ToXin (RTX) cytotoxins. HlyA preferentially binds the ?(L)?(2)integrin LFA-1 (CD11a/CD18) of leukocytes and can promiscuously bind and also permeabilize many other cells. CyaA bears an N-terminal adenylyl cyclase (AC) domain linked to a pore-forming RTX cytolysin (Hly) moiety, binds the complement receptor 3 (CR3, ?(M)?(2), CD11b/CD18, or Mac-1) of myeloid phagocytes, penetrates their plasma membrane, and delivers the AC enzyme into the cytosol. We constructed a set of CyaA/HlyA chimeras and show that the CyaC-acylated segment and the CR3-binding RTX domain of CyaA can be functionally replaced by the HlyC-acylated segment and the much shorter RTX domain of HlyA. Instead of binding CR3, a CyaA(1-710)/HlyA(411-1024)chimera bound the LFA-1 receptor and effectively delivered AC into Jurkat T cells. At high chimera concentrations (25 nm), the interaction with LFA-1 was not required for CyaA(1-710)/HlyA(411-1024)binding to CHO cells. However, interaction with the LFA-1 receptor strongly enhanced the specific capacity of the bound CyaA(1-710)/HlyA(411-1024)chimera to penetrate cells and deliver the AC enzyme into their cytosol. Hence, interaction of the acylated segment and/or the RTX domain of HlyA with LFA-1 promoted a productive membrane interaction of the chimera. These results help delimit residues 400?710 of CyaA as an ?AC translocon? sufficient for translocation of the AC polypeptide across the plasma membrane of target cells.
Permanent Link: http://hdl.handle.net/11104/0311527