Isolation and characterization of the gene encoding single-stranded-DNA-binding protein (SSB) from four marine Shewanella strains that differ in their temperature and pressure optima for growth
The ssb gene, coding for single-stranded-DNA-binding protein (SSB), was cloned from four marine Shewanella strains that differed in their temperature and pressure optima and ranges of growth. All four Shewanellassb genes complemented Escherichia coli ssb point and deletion mutants, with efficiencies that varied with temperature and ssb gene source. The Shewanella SSBs are the largest bacterial SSBs identified to date (24.9-26.3 kDa) and may be divided into conserved amino- and carboxy-terminal regions and a highly variable central region. Greater amino acid sequence homology was observed between the Shewanella SSBs as a group (72-87%) than with other bacterial SSBs (52-69%). Analysis of the amino acid composition of the Shewanella SSBs revealed several features that could correlate with pressure or temperature adaptation. SSBs from the three low-temperature-adapted Shewanella strains were an order of magnitude more hydrophilic than that from the mesophilic strain, and differences in the distribution of eight amino acids were identified which could contribute to either the temperature or pressure adaptation of the proteins. The SSBs from all four Shewanella strains were overproduced and partially purified based upon their ability to bind single-stranded DNA. The differences found among the Shewanella SSBs suggest that these proteins will provide a useful system for exploring the adaptation of protein-protein and protein-DNA interactions at low temperature and high pressure.
BrandsmaJ. A.,
BoschD.,
de RuyterM.,
van de PutteP.1985; Analysis of the regulatory region of the ssb gene of Escherichia coli.
. Nucleic Acids Res 13:5095–5109
CurthU,
Genschelj,
UrbankeC,
GreipelJ.
1996; In vitro and in vivo function of the C-terminus of Escherichia coli singlestranded DNA binding protein. . Nucleic Acids Res 24:2706–2711
DeLongE. F.,
YayanosA. A.1986; Biochemical function and ecological significance of novel bacterial lipids in deep-sea procaryotes. . Appl Environ Microbiol 51:730–737
FoguelD.,
SilvaJ.1994; Cold denaturation of a repressoroperator complex : the role of entropy in protein-DNA recognition. . Proc Natl Acad Sci USA 91:8244–8247
GrossM.,
JaenickeR.1994; Proteins under pressure- the influence of high hydrostatic pressure on structure, function and assembly of protein complexes. . Eur J Biochem 221:617–630
HammarM.,
ArnqvistA.,
BianZ.,
OlsenA.,
NormarkS.1995; Expression of two csg operons is required for production of fibronectin- and Congo red-binding curli polymers in Escherichia coli K-12. . Mol Microbiol 18:661–670
HostomskiZ.,
AppeltK.,
OgdenR. C.1989; High level expression of self-processed HIV-I protease in E. coli using a synthetic gene. . Biochem Biophys Res Commun 161:1056–1063
JensenM. J.,
TeboB. M.,
BaumannP.,
MandelM.,
NealsonK.H.1980; Characterization of Alteromonas hanedai (sp. nov.), a nonfermentative luminous species of marine origin.. Curr Microbiol 3:311–315
LohmanT. M.,
GreenM. J.,
BeyerR. S.1986; Large-scale overproduction and rapid purification of the Escherichia coli ssb gene product. Expression of the ssb gene under lambda PL control.. Biochemistry 25:21–25
MerrillB. M.,
WilliamsK. R.,
ChaseJ. W.,
KonigsbergW. H.1984; Photochemical cross-linking of the Escherichia coli singlestranded DNA-binding protein to oligo deoxynucleotides. Identification of phenylalanine 60 as the site of cross-linking.. J Biol Chem 259:10850–10856
MrabetN. T.,
Van den BroeckA.,
Van den BrandeI.,
13 other authors.
1992; Arginine residues as stabilizing elements in proteins.. Biochemistry 31:2239–2253
PorterR. D.,
BlackS.1991; The single-stranded-DNA-binding protein encoded by the Escherichia coli F factor can complement a deletion of the chromosomal ssb gene.. J Bacteriol 173:2720–2723
PorterR. D.,
BlackS.,
PannuriS.,
CarlsonS.1990; Use of the Escherichia coli ssb gene to prevent bioreactor takeover by plasmidless cells.. Bio/Technology 8:47–51
Rentier-DelrueF.,
MandeS. C.,
MoyensS.,
TerpstraP.,
MainfroidV.,
GorajK.,
LionM.,
HolW. G. J.,
MartialJ. A.1993; Cloning and overexpression of the triosephosphate isomerase genes from psychrophilic and thermophilic bacteria.. J Mol Biol 229:85–93
RoyerC. A.,
ChakerianA. E.,
MathewsK. S.1990; Macromolecular binding equilibria in the lac repressor system : studies using high pressure fluorescence spectroscopy.. Biochem 29:4959–4966
RoyerC. A.,
HinckA. P.,
LohS. N.,
PrehodaK. E.,
PengX.,
JonasJ.,
MarkleyJ. L.1993; Effects of amino acid substitutions
on the pressure denaturation of staphylococcal nuclease as monitored by fluorescence and nuclear magnetic resonance
spectroscopy.. Biochemistry 32:5222–5232
SaikiR. K.,
GelfandD. H.,
StoffelS.,
ScharfS. J.,
HiguchiR.,
HornG. T.,
Mullisk. B,
EhrlichH. A.1998; Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase.. Science 239:487–491
TaborS.,
RichardsonC. C.1985; A bacteriophage T7 RNA polymerase/promoter system for the exclusive expression of specific genes.. Proc Natl Acad Sci USA 82:1074–1078
TobalG. M.1993; Purification and characterization of a malate dehydrogenase from the marine bacterium ShewanellaSC2A. Masters thesis, University of California, San Diego..
de VriesJ.,
WackernagelW.1993; Cloning and sequencing of the Serratia marcescens gene encoding a single-stranded DNAbinding protein (SSB) and its promoter region.. Gene 127:39–45
de VriesJ.,
WackernagelW.1994; Cloning and sequencing of the Proteus mirabilis gene for a single-stranded DNA-binding protein (SSB) and complementation of Escherichia coli ssb point and deletion mutations.. Microbiology 140:889–895
de VriesJ.,
GenschelJ.,
UrbankeC.,
TholeH.,
WackernagelW. The single-stranded-DNA-binding proteins (SSB) of Proteus mirabilis and Serratia marcescens.. Eur J Biochem 224:613–622
YayanosA. A.,
DietzA. S.,
Van BoxtelR.1982; Dependence of reproduction rate on pressure as a hallmark of deep-sea bacteria.. Appl Environ Microbiol 44:1356–1361
ZwicklP.,
FabryS.,
BogedainC.,
HaasA.,
HenselR.1990; Glyceraldehyde-3-phosphate dehydrogenase from the hyper- Received 9 August 1996; revised 24 October 1996; accepted 6 November thermophilic archaebacterium Pyrococcus woesi: characteri-zation of the enzyme, cloning and sequencing of the gene, and expression in Escherichia coli.. J Bacteriof 172:4329–4338
Isolation and characterization of the gene encoding single-stranded-DNA-binding protein (SSB) from four marine Shewanella strains that differ in their temperature and pressure optima for growth