PMID- 32393579
OWN - NLM
STAT- MEDLINE
DCOM- 20210113
LR  - 20210711
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 295
IP  - 28
DP  - 2020 Jul 10
TI  - Retargeting from the CR3 to the LFA-1 receptor uncovers the adenylyl cyclase 
      enzyme-translocating segment of Bordetella adenylate cyclase toxin.
PG  - 9349-9365
LID - 10.1074/jbc.RA120.013630 [doi]
AB  - The Bordetella adenylate cyclase toxin-hemolysin (CyaA) and the alpha-hemolysin 
      (HlyA) of Escherichia coli belong to the family of cytolytic pore-forming Repeats 
      in ToXin (RTX) cytotoxins. HlyA preferentially binds the alpha(L)beta(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, 
      alpha(M)beta(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.
CI  - (c) 2020 Masin et al.
FAU - Masin, Jiri
AU  - Masin J
AD  - Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech 
      Republic masin@biomed.cas.cz osicka@biomed.cas.cz.
FAU - Osickova, Adriana
AU  - Osickova A
AD  - Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech 
      Republic.
AD  - Faculty of Science, Charles University in Prague, Prague, Czech Republic.
FAU - Jurnecka, David
AU  - Jurnecka D
AUID- ORCID: 0000-0002-1690-8908
AD  - Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech 
      Republic.
AD  - Faculty of Science, Charles University in Prague, Prague, Czech Republic.
FAU - Klimova, Nela
AU  - Klimova N
AUID- ORCID: 0000-0002-9425-6386
AD  - Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech 
      Republic.
AD  - Faculty of Science, Charles University in Prague, Prague, Czech Republic.
FAU - Khaliq, Humaira
AU  - Khaliq H
AD  - Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech 
      Republic.
FAU - Sebo, Peter
AU  - Sebo P
AUID- ORCID: 0000-0002-9755-7715
AD  - Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech 
      Republic.
FAU - Osicka, Radim
AU  - Osicka R
AUID- ORCID: 0000-0003-2626-5456
AD  - Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech 
      Republic masin@biomed.cas.cz osicka@biomed.cas.cz.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200511
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Adenylate Cyclase Toxin)
RN  - 0 (Lymphocyte Function-Associated Antigen-1)
RN  - 0 (Macrophage-1 Antigen)
SB  - IM
MH  - Adenylate Cyclase Toxin/*metabolism
MH  - Animals
MH  - *Bordetella
MH  - CHO Cells
MH  - Cricetulus
MH  - Cytosol/*metabolism
MH  - Female
MH  - Humans
MH  - Jurkat Cells
MH  - Lymphocyte Function-Associated Antigen-1/*metabolism
MH  - Macrophage-1 Antigen/*metabolism
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Protein Transport
MH  - THP-1 Cells
PMC - PMC7363143
OTO - NOTNLM
OT  - AC domain translocation
OT  - AC translocon
OT  - Bordetella pertussis
OT  - CyaA
OT  - Escherichia coli (E. coli)
OT  - HlyA
OT  - RTX toxin
OT  - acylation
OT  - acyltransferase
OT  - bacterial toxin
OT  - complement receptor 3 (CR3,)
OT  - fatty acid
OT  - fatty acyl
OT  - integrin
OT  - protein acylation
OT  - protein translocation
COIS- Conflict of interest-The authors declare no conflicts of interest in regards to 
      this manuscript.
EDAT- 2020/05/13 06:00
MHDA- 2021/01/14 06:00
PMCR- 2021/07/10
CRDT- 2020/05/13 06:00
PHST- 2020/03/27 00:00 [received]
PHST- 2020/05/07 00:00 [revised]
PHST- 2020/05/13 06:00 [pubmed]
PHST- 2021/01/14 06:00 [medline]
PHST- 2020/05/13 06:00 [entrez]
PHST- 2021/07/10 00:00 [pmc-release]
AID - S0021-9258(17)48958-7 [pii]
AID - RA120.013630 [pii]
AID - 10.1074/jbc.RA120.013630 [doi]
PST - ppublish
SO  - J Biol Chem. 2020 Jul 10;295(28):9349-9365. doi: 10.1074/jbc.RA120.013630. Epub 
      2020 May 11.