TY - JOUR
T1 - The N-end rule pathway as a nitric oxide sensor controlling the levels of multiple regulators
AU - Hu, Rong Gui
AU - Sheng, Jun
AU - Qi, Xin
AU - Xu, Zhenming
AU - Takahashi, Terry T.
AU - Varshavsky, Alexander
N1 - Funding Information:
Acknowledgements We thank G. Enikolopov, T. Michurina and J. M. Encinas for NOS12/2 mice; A. Mülsch for DNIC-[GSH]2; M. Shahgholi and J. Zhou for MS analyses of peptides; F. Rusnak and G. Hathaway for protein sequencing; J. Racs and S. Horvath for peptide synthesis; L. del Carmen Sandoval, B. W. Kennedy and S. Pease for advice and assistance with mouse mutants; Y. T. Kwon for [UBR12/2UBR22/2] cells; R. Baker for plasmids that enabled the USP2-based ubiquitin fusion technique; G. Eriani and F. Du for gifts of other plasmids; Z. Xia for USP2 enzyme; R. Roberts for use of his laboratory equipment; and E. Graciet and C. Brower for comments on the manuscript. Purification of mouse ATE1-1 was performed at CalTech’s Protein Expression Center by I. K. Nangiana and the late P. Snow. We dedicate this paper to the memory of Dr Snow. This work was supported by grants from the NIH and the Ellison Medical Foundation to A.V.
PY - 2005/10/13
Y1 - 2005/10/13
N2 - The conjugation of arginine to proteins is a part of the N-end rule pathway of protein degradation. Three amino (N)-terminal residues - aspartate, glutamate and cysteine - are arginylated by ATE1-encoded arginyl-transferases. Here we report that oxidation of N-terminal cysteine is essential for its arginylation. The in vivo oxidation of N-terminal cysteine, before its arginylation, is shown to require nitric oxide. We reconstituted this process in vitro as well. The levels of regulatory proteins bearing N-terminal cysteine, such as RGS4, RGS5 and RGS16, are greatly increased in mouse ATE1-/- embryos, which lack arginylation. Stabilization of these proteins, the first physiological substrates of mammalian N-end rule pathway, may underlie cardiovascular defects in ATE1-/- embryos. Our findings identify the N-end rule pathway as a new nitric oxide sensor that functions through its ability to destroy specific regulatory proteins bearing N-terminal cysteine, at rates controlled by nitric oxide and apparently by oxygen as well.
AB - The conjugation of arginine to proteins is a part of the N-end rule pathway of protein degradation. Three amino (N)-terminal residues - aspartate, glutamate and cysteine - are arginylated by ATE1-encoded arginyl-transferases. Here we report that oxidation of N-terminal cysteine is essential for its arginylation. The in vivo oxidation of N-terminal cysteine, before its arginylation, is shown to require nitric oxide. We reconstituted this process in vitro as well. The levels of regulatory proteins bearing N-terminal cysteine, such as RGS4, RGS5 and RGS16, are greatly increased in mouse ATE1-/- embryos, which lack arginylation. Stabilization of these proteins, the first physiological substrates of mammalian N-end rule pathway, may underlie cardiovascular defects in ATE1-/- embryos. Our findings identify the N-end rule pathway as a new nitric oxide sensor that functions through its ability to destroy specific regulatory proteins bearing N-terminal cysteine, at rates controlled by nitric oxide and apparently by oxygen as well.
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U2 - 10.1038/nature04027
DO - 10.1038/nature04027
M3 - Article
C2 - 16222293
AN - SCOPUS:27144557281
SN - 0028-0836
VL - 437
SP - 981
EP - 986
JO - Nature
JF - Nature
IS - 7061
ER -