TY - JOUR

T1 - Evidence against the double-arginine motif as the only determinant for protein translocation by a novel Sec-independent pathway in Escherichia coli

AU - Brüser, Thomas

AU - Deutzmann, Rainer

AU - Dahl, Christiane

N1 - Funding Information: We thank H.-G. Sahl and M. Becker (Bonn, Germany) for production of antiserum. We also thank W. Klipp (Bochum, Germany), A. Seidler (Bochum, Germany) and J. Packer (Cambridge, England) for helpful comments and discussions. We are grateful to H.G. Trüper for his support of the project. T.B. thanks the Cusanuswerk for a grant. Copyright: Copyright 2007 Elsevier B.V., All rights reserved.

PY - 1998/7/15

Y1 - 1998/7/15

N2 - Proteins which are synthesized with a signal peptide containing a 'double-arginine' motif may be translocated across the bacterial cytoplasmic membrane by a mechanism that is different from the known Sec and signal recognition particle pathways. The function of the double-arginine motif as a determinant for this novel pathway was studied by expressions of gene constructs coding for the high potential iron-sulfur protein (HiPIP) from Chromatium vinosum D in Escherichia coli. When the protein was produced with its original double-arginine motif-containing signal peptide, it was in part translocated into the periplasm and thereby processed, as shown by immunoblots after cell fractionation and N-terminal sequencing of purified HiPIP. Processing was not inhibited significantly by 3 mM sodium azide, indicating that translocation of HiPIP occurs by a SecA-independent pathway. Translocation of HiPIP could be altered to the SecA-dependent mode when its signal peptide was substituted by that of PelB from Erwinia carotovora. When the HiPIP double-arginine motif (SRRDAVK) was introduced into the corresponding position of the PelB signal peptide, the transport pathway remained SecA-dependent. This indicates that additional determinants are required for translocation by the Sec-independent pathway. Copyright (C) 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

AB - Proteins which are synthesized with a signal peptide containing a 'double-arginine' motif may be translocated across the bacterial cytoplasmic membrane by a mechanism that is different from the known Sec and signal recognition particle pathways. The function of the double-arginine motif as a determinant for this novel pathway was studied by expressions of gene constructs coding for the high potential iron-sulfur protein (HiPIP) from Chromatium vinosum D in Escherichia coli. When the protein was produced with its original double-arginine motif-containing signal peptide, it was in part translocated into the periplasm and thereby processed, as shown by immunoblots after cell fractionation and N-terminal sequencing of purified HiPIP. Processing was not inhibited significantly by 3 mM sodium azide, indicating that translocation of HiPIP occurs by a SecA-independent pathway. Translocation of HiPIP could be altered to the SecA-dependent mode when its signal peptide was substituted by that of PelB from Erwinia carotovora. When the HiPIP double-arginine motif (SRRDAVK) was introduced into the corresponding position of the PelB signal peptide, the transport pathway remained SecA-dependent. This indicates that additional determinants are required for translocation by the Sec-independent pathway. Copyright (C) 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

KW - Chromatium vinosum

KW - Double-arginine motif signal peptide

KW - Escherichia coli

KW - High potential iron-sulfur protein

KW - Iron-sulfur cluster

KW - Protein translocation

UR - http://www.scopus.com/inward/record.url?scp=0039351902&partnerID=8YFLogxK

U2 - 10.1016/S0378-1097(98)00233-X

DO - 10.1016/S0378-1097(98)00233-X

M3 - Article

C2 - 9682482

AN - SCOPUS:0039351902

VL - 164

SP - 329

EP - 336

JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

SN - 0378-1097

IS - 2

ER -