Two-step Secretion of the Serratia Marcescens Extracellular Nuclease

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1996 Jul 1

PMID 8682779

Citations 3

Authors

Y Suh

S Jin

T K Ball

M J Benedik

Affiliations

Soon will be listed here.

Abstract

The extracellular nuclease of Serratia marcescens is one of a wide variety of enzymes secreted into the growth medium. Its appearance occurs late in the growth of a culture, and its gene, nucA, is transcriptionally regulated in a complex fashion by growth phase and other factors. Pulse-labeling studies reveal that extracellular secretion of nuclease occurs as a two-step process. In the first step, nuclease is rapidly translocated across the cytoplasmic membrane into the periplasm, where it accumulates as a mature active nuclease. A precursor protein, nuclease still carrying its signal sequence, was detected in the presence of carbonyl cyanide m-chlorophenylhydrazone or sodium azide, suggesting that this initial translocation and signal processing step involves an energy-dependent and Sec-dependent pathway in S. marcescens. The second step of secretion across the outer membrane is a slow process requiring between 30 to 120 min, depending on growth conditions.

Citing Articles

Expression and Characterization of Recombinant Serratia liquefaciens Nucleases Produced with Baculovirus-mediated Silkworm Expression System.

Iiyama K, Lee J, Tatsuke T, Mon H, Kusakabe T Mol Biotechnol. 2016; 58(6):393-403.

PMID: 27059494 DOI: 10.1007/s12033-016-9937-y.

Nuclease overexpression mutants of Serratia marcescens.

Guynn L, Dai W, Benedik M J Bacteriol. 1998; 180(8):2262-4.

PMID: 9555916 PMC: 107160. DOI: 10.1128/JB.180.8.2262-2264.1998.

Secretion of nuclease across the outer membrane of Serratia marcescens and its energy requirements.

Suh Y, Benedik M J Bacteriol. 1997; 179(3):677-83.

PMID: 9006020 PMC: 178747. DOI: 10.1128/jb.179.3.677-683.1997.

References

Dums F, Dow J, Daniels M . Structural characterization of protein secretion genes of the bacterial phytopathogen Xanthomonas campestris pathovar campestris: relatedness to secretion systems of other gram-negative bacteria. Mol Gen Genet. 1991; 229(3):357-64. DOI: 10.1007/BF00267456. View

Jones J, Grady K, Suslow T, Bedbrook J . Isolation and characterization of genes encoding two chitinase enzymes from Serratia marcescens. EMBO J. 1986; 5(3):467-73. PMC: 1166787. DOI: 10.1002/j.1460-2075.1986.tb04235.x. View

Nakahama K, Yoshimura K, Marumoto R, Kikuchi M, Lee I, Hase T . Cloning and sequencing of Serratia protease gene. Nucleic Acids Res. 1986; 14(14):5843-55. PMC: 311595. DOI: 10.1093/nar/14.14.5843. View

Heller K . Lipolytic activity copurified with the outer membrane of Serratia marcescens. J Bacteriol. 1979; 140(3):1120-2. PMC: 216763. DOI: 10.1128/jb.140.3.1120-1122.1979. View

Winkler U, Timmis K . Pleiotropic mutations in Serratia marcescens which increase the synthesis of certain exocellular proteins and the rate of spontaneous prophage induction. Mol Gen Genet. 1973; 124(3):197-206. DOI: 10.1007/BF00293091. View

Chen Y, Shipley G, Ball T, Benedik M . Regulatory mutants and transcriptional control of the Serratia marcescens extracellular nuclease gene. Mol Microbiol. 1992; 6(5):643-51. DOI: 10.1111/j.1365-2958.1992.tb01512.x. View

Miller M, Tanner J, Alpaugh M, Benedik M, Krause K . 2.1 A structure of Serratia endonuclease suggests a mechanism for binding to double-stranded DNA. Nat Struct Biol. 1994; 1(7):461-8. DOI: 10.1038/nsb0794-461. View

Givskov M, Olsen L, Molin S . Cloning and expression in Escherichia coli of the gene for extracellular phospholipase A1 from Serratia liquefaciens. J Bacteriol. 1988; 170(12):5855-62. PMC: 211692. DOI: 10.1128/jb.170.12.5855-5862.1988. View

Akatsuka H, Kawai E, Omori K, Komatsubara S, Shibatani T, Tosa T . The lipA gene of Serratia marcescens which encodes an extracellular lipase having no N-terminal signal peptide. J Bacteriol. 1994; 176(7):1949-56. PMC: 205299. DOI: 10.1128/jb.176.7.1949-1956.1994. View

10.

Ball T, Saurugger P, Benedik M . The extracellular nuclease gene of Serratia marcescens and its secretion from Escherichia coli. Gene. 1987; 57(2-3):183-92. DOI: 10.1016/0378-1119(87)90121-1. View

11.

Oliver D, Cabelli R, Dolan K, Jarosik G . Azide-resistant mutants of Escherichia coli alter the SecA protein, an azide-sensitive component of the protein export machinery. Proc Natl Acad Sci U S A. 1990; 87(21):8227-31. PMC: 54928. DOI: 10.1073/pnas.87.21.8227. View

12.

Reeves P, Whitcombe D, Wharam S, Gibson M, Allison G, Bunce N . Molecular cloning and characterization of 13 out genes from Erwinia carotovora subspecies carotovora: genes encoding members of a general secretion pathway (GSP) widespread in gram-negative bacteria. Mol Microbiol. 1993; 8(3):443-56. DOI: 10.1111/j.1365-2958.1993.tb01589.x. View

13.

Ball T, Suh Y, Benedik M . Disulfide bonds are required for Serratia marcescens nuclease activity. Nucleic Acids Res. 1992; 20(19):4971-4. PMC: 334271. DOI: 10.1093/nar/20.19.4971. View

14.

Koronakis V, Hughes C, Koronakis E . Energetically distinct early and late stages of HlyB/HlyD-dependent secretion across both Escherichia coli membranes. EMBO J. 1991; 10(11):3263-72. PMC: 453051. DOI: 10.1002/j.1460-2075.1991.tb04890.x. View

15.

Jin S, Chen Y, Christie G, Benedik M . Regulation of the Serratia marcescens extracellular nuclease: positive control by a homolog of P2 Ogr encoded by a cryptic prophage. J Mol Biol. 1996; 256(2):264-78. DOI: 10.1006/jmbi.1996.0084. View

16.

Lindeberg M, Collmer A . Analysis of eight out genes in a cluster required for pectic enzyme secretion by Erwinia chrysanthemi: sequence comparison with secretion genes from other gram-negative bacteria. J Bacteriol. 1992; 174(22):7385-97. PMC: 207435. DOI: 10.1128/jb.174.22.7385-7397.1992. View

17.

Kunkel T, Roberts J, Zakour R . Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987; 154:367-82. DOI: 10.1016/0076-6879(87)54085-x. View

18.

Salmond G, Reeves P . Membrane traffic wardens and protein secretion in gram-negative bacteria. Trends Biochem Sci. 1993; 18(1):7-12. DOI: 10.1016/0968-0004(93)90080-7. View

19.

Spratt B . A vector for the construction of translational fusions to TEM beta-lactamase and the analysis of protein export signals and membrane protein topology. Gene. 1986; 49(3):341-9. DOI: 10.1016/0378-1119(86)90370-7. View

20.

Filloux A, Bally M, Ball G, Akrim M, Tommassen J, Lazdunski A . Protein secretion in gram-negative bacteria: transport across the outer membrane involves common mechanisms in different bacteria. EMBO J. 1990; 9(13):4323-9. PMC: 552217. DOI: 10.1002/j.1460-2075.1990.tb07881.x. View