:The short-term goal of this project is to understand the trafficking and activation of pertussis toxin (PT) within mammalian cells. Bordetella pertussis colonizes the human respiratory tract and causes the disease pertussis (whooping cough). Several systemic symptoms accompany this disease, even though B. pertussis is not thought to invade beyond the respiratory tract. Instead these symptoms are thought to be due to the action of PT, an exotoxin produced by B. pertussis. PT is an ADP ribosyltransferase that modifies several heterotrimeric G proteins, causing a wide range of effects on signaling in mammalian cells. How PT is transported within mammalian cells to arrive at its target proteins in an active form is largely unknown. In addition, how PT is transported from the respiratory tract to systemic sites is completely unknown. Understanding the mechanisms utilized by this complex toxin to achieve these effects will provide key information on the cell biology of PT and will help to provide a groundwork for studies to elucidate the role of this toxin in B. pertussis infection and disease. In addition, this information may allow development of therapeutics to combat the effects of the toxin and may also allow improvement of existing pertussis vaccines that include PT or development of novel vaccine molecules using PT as a intracellular delivery vector. We have preliminary data indicating that (i) PT may undergo retrograde intracellular trafficking through the Golgi apparatus and endoplasmic reticulum (ER) en route to its cytosolic target proteins, (ii) that proteolytic processing of the active Si subunit of cell-associated PT occurs and may be important for its activity, and (iii) that there is apparent transcytosis of active PT across intact polarized epithelial cells in culture. Therefore the specific aims of this proposal are to investigate the trafficking, processing and transcytosis of PT in mammalian cells and the key features of this toxin that mediate these events.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI050022-03
Application #
6632480
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Program Officer
Klein, David L
Project Start
2001-07-01
Project End
2006-05-31
Budget Start
2003-06-01
Budget End
2004-05-31
Support Year
3
Fiscal Year
2003
Total Cost
$297,000
Indirect Cost
Name
University of Maryland Baltimore
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
188435911
City
Baltimore
State
MD
Country
United States
Zip Code
21201
Plaut, Roger D; Scanlon, Karen M; Taylor, Michael et al. (2016) Intracellular disassembly and activity of pertussis toxin require interaction with ATP. Pathog Dis 74:
Pascual, Julio; Samaniego, Milagros D; Torrealba, Jose R et al. (2008) Antibody-mediated rejection of the kidney after simultaneous pancreas-kidney transplantation. J Am Soc Nephrol 19:812-24
Plaut, Roger D; Carbonetti, Nicholas H (2008) Retrograde transport of pertussis toxin in the mammalian cell. Cell Microbiol 10:1130-9
Carbonetti, Nicholas H; Artamonova, Galina V; Van Rooijen, Nico et al. (2007) Pertussis toxin targets airway macrophages to promote Bordetella pertussis infection of the respiratory tract. Infect Immun 75:1713-20
Worthington, Zoe E V; Carbonetti, Nicholas H (2007) Evading the proteasome: absence of lysine residues contributes to pertussis toxin activity by evasion of proteasome degradation. Infect Immun 75:2946-53
Carbonetti, Nicholas H (2007) Immunomodulation in the pathogenesis of Bordetella pertussis infection and disease. Curr Opin Pharmacol 7:272-8
Carbonetti, Nicholas H; Mays, R Michael; Artamonova, Galina V et al. (2005) Proteolytic cleavage of pertussis toxin S1 subunit is not essential for its activity in mammalian cells. BMC Microbiol 5:7
Carbonetti, Nicholas H; Artamonova, Galina V; Andreasen, Charlotte et al. (2005) Pertussis toxin and adenylate cyclase toxin provide a one-two punch for establishment of Bordetella pertussis infection of the respiratory tract. Infect Immun 73:2698-703
Carbonetti, Nicholas H; Artamonova, Galina V; Andreasen, Charlotte et al. (2004) Suppression of serum antibody responses by pertussis toxin after respiratory tract colonization by Bordetella pertussis and identification of an immunodominant lipoprotein. Infect Immun 72:3350-8
Carbonetti, Nicholas H; Artamonova, Galina V; Mays, R Michael et al. (2003) Pertussis toxin plays an early role in respiratory tract colonization by Bordetella pertussis. Infect Immun 71:6358-66