PrlA4 Prevents the Rejection of Signal Sequence Defective Preproteins by Stabilizing the SecA-SecY Interaction During the Initiation of Translocation

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1998 Jul 3

PMID 9649433

Citations 35

Authors

J P van der Wolk

P Fekkes

A Boorsma

J L Huie

T J Silhavy

A J Driessen

Affiliations

Soon will be listed here.

Abstract

In Escherichia coli, precursor proteins are translocated across the cytoplasmic membrane by translocase. This multisubunit enzyme consists of a preprotein-binding and ATPase domain, SecA, and the SecYEG complex as the integral membrane domain. PrlA4 is a mutant of SecY that enables the translocation of preproteins with a defective, or missing, signal sequence. Inner membranes of the prlA4 strain efficiently translocate Delta8proOmpA, a proOmpA derivative with a non-functional signal sequence. Owing to the signal sequence mutation, Delta8proOmpA binds to the translocase with a lowered affinity and the recognition is not restored by the prlA4 SecY. At the ATP-dependent initiation of translocation, the binding affinity of SecA for SecYEG is lowered causing the premature loss of bound preproteins from the translocase. The prlA4 membranes, however, bind SecA with a much higher affinity than the wild-type, and during initiation, the SecA and preprotein remain bound at the translocation site allowing an improved efficiency of translocation. It is concluded that the prlA4 strain prevents the rejection of defective preproteins from the export pathway by stabilizing SecA at the SecYEG complex.

Citing Articles

Strategies to Enhance Periplasmic Recombinant Protein Production Yields in .

Karyolaimos A, Gier J Front Bioeng Biotechnol. 2021; 9:797334.

PMID: 34970535 PMC: 8712718. DOI: 10.3389/fbioe.2021.797334.

SecY-mediated quality control prevents the translocation of non-gated porins.

Jung S, Bader V, Natriashvili A, Koch H, Winklhofer K, Tatzelt J Sci Rep. 2020; 10(1):16347.

PMID: 33004891 PMC: 7530735. DOI: 10.1038/s41598-020-73185-y.

Comparison of Single and Multiple Turnovers of SecYEG in Escherichia coli.

Mao C, Bariya P, Suo Y, Randall L J Bacteriol. 2020; 202(24).

PMID: 32989086 PMC: 7685557. DOI: 10.1128/JB.00462-20.

SecA-Mediated Protein Translocation through the SecYEG Channel.

Komarudin A, Driessen A Microbiol Spectr. 2019; 7(4).

PMID: 31373268 PMC: 10957188. DOI: 10.1128/microbiolspec.PSIB-0028-2019.

The Sec System: Protein Export in .

Crane J, Randall L EcoSal Plus. 2017; 7(2).

PMID: 29165233 PMC: 5807066. DOI: 10.1128/ecosalplus.ESP-0002-2017.

References

Crooke E, Wickner W . Trigger factor: a soluble protein that folds pro-OmpA into a membrane-assembly-competent form. Proc Natl Acad Sci U S A. 1987; 84(15):5216-20. PMC: 298825. DOI: 10.1073/pnas.84.15.5216. View

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

Cabelli R, Chen L, Tai P, Oliver D . SecA protein is required for secretory protein translocation into E. coli membrane vesicles. Cell. 1988; 55(4):683-92. DOI: 10.1016/0092-8674(88)90227-9. View

Sako T, Iino T . Distinct mutation sites in prlA suppressor mutant strains of Escherichia coli respond either to suppression of signal peptide mutations or to blockage of staphylokinase processing. J Bacteriol. 1988; 170(11):5389-91. PMC: 211621. DOI: 10.1128/jb.170.11.5389-5391.1988. View

Weiss J, Ray P, Bassford Jr P . Purified secB protein of Escherichia coli retards folding and promotes membrane translocation of the maltose-binding protein in vitro. Proc Natl Acad Sci U S A. 1988; 85(23):8978-82. PMC: 282643. DOI: 10.1073/pnas.85.23.8978. View

Fikes J, Bassford Jr P . Novel secA alleles improve export of maltose-binding protein synthesized with a defective signal peptide. J Bacteriol. 1989; 171(1):402-9. PMC: 209602. DOI: 10.1128/jb.171.1.402-409.1989. View

Cunningham K, Lill R, Crooke E, Rice M, Moore K, Wickner W . SecA protein, a peripheral protein of the Escherichia coli plasma membrane, is essential for the functional binding and translocation of proOmpA. EMBO J. 1989; 8(3):955-9. PMC: 400896. DOI: 10.1002/j.1460-2075.1989.tb03457.x. View

Lill R, Cunningham K, Brundage L, Ito K, Oliver D, Wickner W . SecA protein hydrolyzes ATP and is an essential component of the protein translocation ATPase of Escherichia coli. EMBO J. 1989; 8(3):961-6. PMC: 400897. DOI: 10.1002/j.1460-2075.1989.tb03458.x. View

Stader J, Gansheroff L, Silhavy T . New suppressors of signal-sequence mutations, prlG, are linked tightly to the secE gene of Escherichia coli. Genes Dev. 1989; 3(7):1045-52. DOI: 10.1101/gad.3.7.1045. View

10.

Yamada H, Matsuyama S, Tokuda H, Mizushima S . A high concentration of SecA allows proton motive force-independent translocation of a model secretory protein into Escherichia coli membrane vesicles. J Biol Chem. 1989; 264(31):18577-81. View

11.

Cunningham K, Wickner W . Specific recognition of the leader region of precursor proteins is required for the activation of translocation ATPase of Escherichia coli. Proc Natl Acad Sci U S A. 1989; 86(22):8630-4. PMC: 298341. DOI: 10.1073/pnas.86.22.8630. View

12.

Lill R, Dowhan W, Wickner W . The ATPase activity of SecA is regulated by acidic phospholipids, SecY, and the leader and mature domains of precursor proteins. Cell. 1990; 60(2):271-80. DOI: 10.1016/0092-8674(90)90742-w. View

13.

Lecker S, Driessen A, Wickner W . ProOmpA contains secondary and tertiary structure prior to translocation and is shielded from aggregation by association with SecB protein. EMBO J. 1990; 9(7):2309-14. PMC: 551957. DOI: 10.1002/j.1460-2075.1990.tb07402.x. View

14.

Brundage L, Hendrick J, Schiebel E, Driessen A, Wickner W . The purified E. coli integral membrane protein SecY/E is sufficient for reconstitution of SecA-dependent precursor protein translocation. Cell. 1990; 62(4):649-57. DOI: 10.1016/0092-8674(90)90111-q. View

15.

Bieker K, Phillips G, Silhavy T . The sec and prl genes of Escherichia coli. J Bioenerg Biomembr. 1990; 22(3):291-310. DOI: 10.1007/BF00763169. View

16.

Hartl F, Lecker S, Schiebel E, Hendrick J, Wickner W . The binding cascade of SecB to SecA to SecY/E mediates preprotein targeting to the E. coli plasma membrane. Cell. 1990; 63(2):269-79. DOI: 10.1016/0092-8674(90)90160-g. View

17.

Shiozuka K, Tani K, Mizushima S, Tokuda H . The proton motive force lowers the level of ATP required for the in vitro translocation of a secretory protein in Escherichia coli. J Biol Chem. 1990; 265(31):18843-7. View

18.

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

19.

Schiebel E, Driessen A, Hartl F, Wickner W . Delta mu H+ and ATP function at different steps of the catalytic cycle of preprotein translocase. Cell. 1991; 64(5):927-39. DOI: 10.1016/0092-8674(91)90317-r. View

20.

Tanji Y, Gennity J, Pollitt S, Inouye M . Effect of OmpA signal peptide mutations on OmpA secretion, synthesis, and assembly. J Bacteriol. 1991; 173(6):1997-2005. PMC: 207733. DOI: 10.1128/jb.173.6.1997-2005.1991. View