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Proc Natl Acad Sci U S A. Dec 1, 1993; 90(23): 11247–11251.
PMCID: PMC47959

A human mitochondrial ATP-dependent protease that is highly homologous to bacterial Lon protease.

Abstract

We have cloned a human ATP-dependent protease that is highly homologous to members of the bacterial Lon protease family. The cloned gene encodes a protein of 963 amino acids with a calculated molecular mass of 106 kDa, slightly higher than that observed by Western blotting the protein from human tissues and cell lines (100 kDa). A single species of mRNA was found for this Lon protease in all human tissues examined. The protease is encoded in the nucleus, and the amino-terminal portion of the protein sequence contains a potential mitochondrial targeting presequence. Immunofluorescence microscopy suggested a predominantly mitochondrial localization for the Lon protease in cultured human cells. A truncated LON gene, in which translation was initiated at Met118 of the coding sequence, was expressed in Escherichia coli and produced a protease that degraded alpha-casein in vitro in an ATP-dependent manner and had other properties similar to E. coli Lon protease.

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  • Gottesman S, Maurizi MR. Regulation by proteolysis: energy-dependent proteases and their targets. Microbiol Rev. 1992 Dec;56(4):592–621. [PMC free article] [PubMed]
  • Goldberg AL. The mechanism and functions of ATP-dependent proteases in bacterial and animal cells. Eur J Biochem. 1992 Jan 15;203(1-2):9–23. [PubMed]
  • Charette MF, Henderson GW, Markovitz A. ATP hydrolysis-dependent protease activity of the lon (capR) protein of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4728–4732. [PMC free article] [PubMed]
  • Chung CH, Goldberg AL. The product of the lon (capR) gene in Escherichia coli is the ATP-dependent protease, protease La. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4931–4935. [PMC free article] [PubMed]
  • Maurizi MR, Trisler P, Gottesman S. Insertional mutagenesis of the lon gene in Escherichia coli: lon is dispensable. J Bacteriol. 1985 Dec;164(3):1124–1135. [PMC free article] [PubMed]
  • Goff SA, Goldberg AL. An increased content of protease La, the lon gene product, increases protein degradation and blocks growth in Escherichia coli. J Biol Chem. 1987 Apr 5;262(10):4508–4515. [PubMed]
  • Gottesman S. Genetics of proteolysis in Escherichia coli*. Annu Rev Genet. 1989;23:163–198. [PubMed]
  • Ito K, Udaka S, Yamagata H. Cloning, characterization, and inactivation of the Bacillus brevis lon gene. J Bacteriol. 1992 Apr;174(7):2281–2287. [PMC free article] [PubMed]
  • Tojo N, Inouye S, Komano T. The lonD gene is homologous to the lon gene encoding an ATP-dependent protease and is essential for the development of Myxococcus xanthus. J Bacteriol. 1993 Jul;175(14):4545–4549. [PMC free article] [PubMed]
  • Tojo N, Inouye S, Komano T. Cloning and nucleotide sequence of the Myxococcus xanthus lon gene: indispensability of lon for vegetative growth. J Bacteriol. 1993 Apr;175(8):2271–2277. [PMC free article] [PubMed]
  • Gill RE, Karlok M, Benton D. Myxococcus xanthus encodes an ATP-dependent protease which is required for developmental gene transcription and intercellular signaling. J Bacteriol. 1993 Jul;175(14):4538–4544. [PMC free article] [PubMed]
  • Katayama-Fujimura Y, Gottesman S, Maurizi MR. A multiple-component, ATP-dependent protease from Escherichia coli. J Biol Chem. 1987 Apr 5;262(10):4477–4485. [PubMed]
  • Hwang BJ, Park WJ, Chung CH, Goldberg AL. Escherichia coli contains a soluble ATP-dependent protease (Ti) distinct from protease La. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5550–5554. [PMC free article] [PubMed]
  • Hershko A, Ciechanover A. The ubiquitin system for protein degradation. Annu Rev Biochem. 1992;61:761–807. [PubMed]
  • Adams MD, Kerlavage AR, Fields C, Venter JC. 3,400 new expressed sequence tags identify diversity of transcripts in human brain. Nat Genet. 1993 Jul;4(3):256–267. [PubMed]
  • Adams MD, Dubnick M, Kerlavage AR, Moreno R, Kelley JM, Utterback TR, Nagle JW, Fields C, Venter JC. Sequence identification of 2,375 human brain genes. Nature. 1992 Feb 13;355(6361):632–634. [PubMed]
  • Gottesman S, Squires C, Pichersky E, Carrington M, Hobbs M, Mattick JS, Dalrymple B, Kuramitsu H, Shiroza T, Foster T, et al. Conservation of the regulatory subunit for the Clp ATP-dependent protease in prokaryotes and eukaryotes. Proc Natl Acad Sci U S A. 1990 May;87(9):3513–3517. [PMC free article] [PubMed]
  • Maurizi MR, Clark WP, Kim SH, Gottesman S. Clp P represents a unique family of serine proteases. J Biol Chem. 1990 Jul 25;265(21):12546–12552. [PubMed]
  • Dubiel W, Ferrell K, Pratt G, Rechsteiner M. Subunit 4 of the 26 S protease is a member of a novel eukaryotic ATPase family. J Biol Chem. 1992 Nov 15;267(32):22699–22702. [PubMed]
  • Klausner RD, Sitia R. Protein degradation in the endoplasmic reticulum. Cell. 1990 Aug 24;62(4):611–614. [PubMed]
  • Desautels M, Goldberg AL. Demonstration of an ATP-dependent, vanadate-sensitive endoprotease in the matrix of rat liver mitochondria. J Biol Chem. 1982 Oct 10;257(19):11673–11679. [PubMed]
  • Watabe S, Kimura T. Adrenal cortex mitochondrial enzyme with ATP-dependent protease and protein-dependent ATPase activities. Purification and properties. J Biol Chem. 1985 Nov 25;260(27):14498–14504. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Studier FW, Rosenberg AH, Dunn JJ, Dubendorff JW. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. [PubMed]
  • Bruggemann EP, Chaudhary V, Gottesman MM, Pastan I. Pseudomonas exotoxin fusion proteins are potent immunogens for raising antibodies against P-glycoprotein. Biotechniques. 1991 Feb;10(2):202–209. [PubMed]
  • Short JM, Fernandez JM, Sorge JA, Huse WD. Lambda ZAP: a bacteriophage lambda expression vector with in vivo excision properties. Nucleic Acids Res. 1988 Aug 11;16(15):7583–7600. [PMC free article] [PubMed]
  • Chin DT, Goff SA, Webster T, Smith T, Goldberg AL. Sequence of the lon gene in Escherichia coli. A heat-shock gene which encodes the ATP-dependent protease La. J Biol Chem. 1988 Aug 25;263(24):11718–11728. [PubMed]
  • Amerik AYu, Antonov VK, Gorbalenya AE, Kotova SA, Rotanova TV, Shimbarevich EV. Site-directed mutagenesis of La protease. A catalytically active serine residue. FEBS Lett. 1991 Aug 5;287(1-2):211–214. [PubMed]
  • Walker JE, Saraste M, Runswick MJ, Gay NJ. Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J. 1982;1(8):945–951. [PMC free article] [PubMed]
  • Maurizi MR. Degradation in vitro of bacteriophage lambda N protein by Lon protease from Escherichia coli. J Biol Chem. 1987 Feb 25;262(6):2696–2703. [PubMed]
  • Waxman L, Goldberg AL. Protease La, the lon gene product, cleaves specific fluorogenic peptides in an ATP-dependent reaction. J Biol Chem. 1985 Oct 5;260(22):12022–12028. [PubMed]
  • Kozak M. Structural features in eukaryotic mRNAs that modulate the initiation of translation. J Biol Chem. 1991 Oct 25;266(30):19867–19870. [PubMed]
  • Gaskins CJ, Smith JF, Ogilvie MK, Hanas JS. Comparison of the sequence and structure of transcription factor IIIA from Bufo americanus and Rana pipiens. Gene. 1992 Oct 21;120(2):197–206. [PubMed]
  • Hendrick JP, Hodges PE, Rosenberg LE. Survey of amino-terminal proteolytic cleavage sites in mitochondrial precursor proteins: leader peptides cleaved by two matrix proteases share a three-amino acid motif. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4056–4060. [PMC free article] [PubMed]
  • von Heijne G, Steppuhn J, Herrmann RG. Domain structure of mitochondrial and chloroplast targeting peptides. Eur J Biochem. 1989 Apr 1;180(3):535–545. [PubMed]
  • Studier FW, Moffatt BA. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. [PubMed]
  • Wallace DC. Diseases of the mitochondrial DNA. Annu Rev Biochem. 1992;61:1175–1212. [PubMed]
  • Desautels M, Goldberg AL. Liver mitochondria contain an ATP-dependent, vanadate-sensitive pathway for the degradation of proteins. Proc Natl Acad Sci U S A. 1982 Mar;79(6):1869–1873. [PMC free article] [PubMed]
  • Glover LA, Lindsay JG. Targeting proteins to mitochondria: a current overview. Biochem J. 1992 Jun 15;284(Pt 3):609–620. [PMC free article] [PubMed]
  • Hahn S, Pinkham J, Wei R, Miller R, Guarente L. The HAP3 regulatory locus of Saccharomyces cerevisiae encodes divergent overlapping transcripts. Mol Cell Biol. 1988 Feb;8(2):655–663. [PMC free article] [PubMed]

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