Recognition, targeting, and hydrolysis of the lambda O replication protein by the ClpP/ClpX protease

Gonciarz-Swiatek, M.; Wawrzynow, Alicja; Um, S. J.; Learn, B. A.; McMacken, R.; Kelley, William; Georgopoulos, C.; Sliekers, O.; Zylicz, M.

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Title		Recognition, targeting, and hydrolysis of the lambda O replication protein by the ClpP/ClpX protease
Authors		Gonciarz-Swiatek, M. Wawrzynow, Alicja Um, S. J. Learn, B. A. McMacken, R. Kelley, William Georgopoulos, C. Sliekers, O. Zylicz, M. show all authors [1 - 9]
Published in		Journal of Biological Chemistry. 1999, vol. 274, no. 20, p. 13999-14005
Abstract		It has previously been established that sequences at the C termini of polypeptide substrates are critical for efficient hydrolysis by the ClpP/ClpX ATP-dependent protease. We report for the bacteriophage lambda O replication protein, however, that N-terminal sequences play the most critical role in facilitating proteolysis by ClpP/ClpX. The N-terminal portion of lambda O is degraded at a rate comparable with that of wild type O protein, whereas the C-terminal domain of O is hydrolyzed at least 10-fold more slowly. Consistent with these results, deletion of the first 18 amino acids of lambda O blocks degradation of the N-terminal domain, whereas proteolysis of the O C-terminal domain is only slightly diminished as a result of deletion of the C-terminal 15 amino acids. We demonstrate that ClpX retains its capacity to bind to the N-terminal domain following removal of the first 18 amino acids of O. However, ClpX cannot efficiently promote the ATP-dependent binding of this truncated O polypeptide to ClpP, the catalytic subunit of the ClpP/ClpX protease. Based on our results with lambda O protein, we suggest that two distinct structural elements may be required in substrate polypeptides to enable efficient hydrolysis by the ClpP/ClpX protease: (i) a ClpX-binding site, which may be located remotely from substrate termini, and (ii) a proper N- or C-terminal sequence, whose exposure on the substrate surface may be induced by the binding of ClpX.
Keywords		Adenosine Triphosphatases/metabolism — Amino Acid Sequence — Bacteriophage lambda/metabolism/physiology — Binding Sites — Endopeptidase Clp — Enzyme-Linked Immunosorbent Assay — Hydrolysis — Kinetics — Molecular Sequence Data — Mutagenesis, Site-Directed — Protein Binding — Serine Endopeptidases/metabolism — Viral Proteins/genetics/metabolism — *Virus Replication
Identifiers		PMID: 10318812
Full text		Article - Limited access to UNIGE Other version: http://www.jbc.org/content/274/20/13999.full.pdf
Structures		Faculté de médecine / Section de médecine clinique / Département de médecine
Citation (ISO format)		GONCIARZ-SWIATEK, M. et al. Recognition, targeting, and hydrolysis of the lambda O replication protein by the ClpP/ClpX protease. In: Journal of Biological Chemistry, 1999, vol. 274, n° 20, p. 13999-14005. https://archive-ouverte.unige.ch/unige:7280