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

Mašín, Jiří; Osičková, Adriana; Jurnečka, David; Klímová, Nela; Khaliq, Humaira; Šebo, Peter; Osička, Radim

doi:https://dx.doi.org/10.1074/jbc.RA120.013630

Number of the records: 1

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

1.

SYSNO ASEP	0533024
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Retargeting from the CR3 to the LFA-1 receptor uncovers the adenylyl cyclase enzyme?translocating segment ofBordetellaadenylate cyclase toxin
Author(s)	Mašín, Jiří (MBU-M)_{RID, ORCID} Osičková, Adriana (MBU-M)_{RID, ORCID} Jurnečka, David (MBU-M)_ORCID Klímová, Nela (MBU-M)_ORCID Khaliq, Humaira (MBU-M) Šebo, Peter (MBU-M)_{RID, ORCID} Osička, Radim (MBU-M)_{RID, ORCID}
Source Title	Journal of Biological Chemistry. - : Elsevier - ISSN 0021-9258 Roč. 295, č. 28 (2020), s. 9349-9365
Number of pages	17 s.
Language	eng - English
Country	US - United States
Keywords	AC domain translocation ; acylation ; acyltransferase ; fatty acyl ; integrin ; RTX toxin
Subject RIV	EE - Microbiology, Virology
OECD category	Microbiology
R&D Projects	GA19-04607S GA ČR - Czech Science Foundation (CSF)
	GA18-18079S GA ČR - Czech Science Foundation (CSF)
	GA19-12695S GA ČR - Czech Science Foundation (CSF)
	GA18-20621S GA ČR - Czech Science Foundation (CSF)
	LM2018133 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
Method of publishing	Open access with time embargo (01.08.2021)
Institutional support	MBU-M - RVO:61388971
UT WOS	000552758600008
EID SCOPUS	85088204783
DOI	10.1074/jbc.RA120.013630
Annotation	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.
Workplace	Institute of Microbiology
Contact	Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231
Year of Publishing	2021
Electronic address	https://www.jbc.org/content/early/2020/05/11/jbc.RA120.013630.short

Number of the records: 1