Characterization of enzymes from Legionella pneumophila involved in reversible adenylylation of Rab1 protein
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Characterization of enzymes from Legionella pneumophila involved in reversible adenylylation of Rab1 protein. / Shkumatov, Alexander V; Oesterlin, Lena K; Schoebel, Stefan; Goody, Philip R; Goody, Roger S; Itzen, Aymelt.
In: J BIOL CHEM, Vol. 287, No. 42, 12.10.2012, p. 35036-46.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Characterization of enzymes from Legionella pneumophila involved in reversible adenylylation of Rab1 protein
AU - Shkumatov, Alexander V
AU - Oesterlin, Lena K
AU - Schoebel, Stefan
AU - Goody, Philip R
AU - Goody, Roger S
AU - Itzen, Aymelt
PY - 2012/10/12
Y1 - 2012/10/12
N2 - After the pathogenic bacterium Legionella pneumophila is phagocytosed, it injects more than 250 different proteins into the cytoplasm of host cells to evade lysosomal digestion and to replicate inside the host cell. Among these secreted proteins is the protein DrrA/SidM, which has been shown to modify Rab1b, a main regulator of vesicular trafficking in eukaryotic cells, by transfer of adenosine monophosphate (AMP) to Tyr(77). In addition, Legionella provides the protein SidD that hydrolytically reverses the covalent modification, suggesting a tight spatial and temporal control of Rab1 function by Legionella during infection. Small angle x-ray scattering experiments of DrrA allowed us to validate a tentative complex model built by combining available crystallographic data. We have established the effects of adenylylation on Rab1 interactions and properties in a quantitative way. In addition, we have characterized the kinetics of DrrA-catalyzed adenylylation as well as SidD-catalyzed deadenylylation toward Rab1 and have determined the nucleotide specificities of both enzymes. This study enhances our knowledge of proteins subverting Rab1 function at the Legionella-containing vacuole.
AB - After the pathogenic bacterium Legionella pneumophila is phagocytosed, it injects more than 250 different proteins into the cytoplasm of host cells to evade lysosomal digestion and to replicate inside the host cell. Among these secreted proteins is the protein DrrA/SidM, which has been shown to modify Rab1b, a main regulator of vesicular trafficking in eukaryotic cells, by transfer of adenosine monophosphate (AMP) to Tyr(77). In addition, Legionella provides the protein SidD that hydrolytically reverses the covalent modification, suggesting a tight spatial and temporal control of Rab1 function by Legionella during infection. Small angle x-ray scattering experiments of DrrA allowed us to validate a tentative complex model built by combining available crystallographic data. We have established the effects of adenylylation on Rab1 interactions and properties in a quantitative way. In addition, we have characterized the kinetics of DrrA-catalyzed adenylylation as well as SidD-catalyzed deadenylylation toward Rab1 and have determined the nucleotide specificities of both enzymes. This study enhances our knowledge of proteins subverting Rab1 function at the Legionella-containing vacuole.
KW - Bacterial Proteins
KW - Guanine Nucleotide Exchange Factors
KW - Humans
KW - Legionella pneumophila
KW - Legionnaires' Disease
KW - Protein Processing, Post-Translational
KW - rab1 GTP-Binding Proteins
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1074/jbc.M112.396861
DO - 10.1074/jbc.M112.396861
M3 - SCORING: Journal article
C2 - 22872634
VL - 287
SP - 35036
EP - 35046
JO - J BIOL CHEM
JF - J BIOL CHEM
SN - 0021-9258
IS - 42
ER -
