Programmed Ribosomal Frameshifting Generates a Copper Transporter and a Copper Chaperone from the Same Gene

Sezen Meydan; Dorota Klepacki; Subbulakshmi Karthikeyan; Tõnu Margus; Paul Thomas; John E. Jones; Yousuf Khan; Joseph Briggs; Jonathan D. Dinman; Nora Vázquez-Laslop; Alexander S. Mankin

doi:10.1016/j.molcel.2016.12.008

Programmed Ribosomal Frameshifting Generates a Copper Transporter and a Copper Chaperone from the Same Gene

Sezen Meydan, Dorota Klepacki, Subbulakshmi Karthikeyan, Tõnu Margus, Paul Thomas, John E. Jones, Yousuf Khan, Joseph Briggs, Jonathan D. Dinman, Nora Vázquez-Laslop^*, Alexander S. Mankin

^*Corresponding author for this work

PCE - Proteomics Center of Excellence

Research output: Contribution to journal › Article › peer-review

70 Scopus citations

Abstract

Metal efflux pumps maintain ion homeostasis in the cell. The functions of the transporters are often supported by chaperone proteins, which scavenge the metal ions from the cytoplasm. Although the copper ion transporter CopA has been known in Escherichia coli, no gene for its chaperone had been identified. We show that the CopA chaperone is expressed in E. coli from the same gene that encodes the transporter. Some ribosomes translating copA undergo programmed frameshifting, terminate translation in the −1 frame, and generate the 70 aa-long polypeptide CopA(Z), which helps cells survive toxic copper concentrations. The high efficiency of frameshifting is achieved by the combined stimulatory action of a “slippery” sequence, an mRNA pseudoknot, and the CopA nascent chain. Similar mRNA elements are not only found in the copA genes of other bacteria but are also present in ATP7B, the human homolog of copA, and direct ribosomal frameshifting in vivo.

Original language	English (US)
Pages (from-to)	207-219
Number of pages	13
Journal	Molecular cell
Volume	65
Issue number	2
DOIs	https://doi.org/10.1016/j.molcel.2016.12.008
State	Published - Jan 19 2017

Funding

We thank Pavel Baranov and John Atkins for helpful advice; Kurt Fredrick and Hyunwoo Lee for providing the pEK4 and pKOV plasmids, respectively; Andrew Jin for help with some experiments; and Dimple Modi, Nikolay Aleksashin, and Hao Lei for advice with some experimental procedures. This work was supported by the grants MCB 1244455 and MCB 1615851 (both to A.S.M. and N.V.-L.) from the National Science Foundation and R01 GM117177 and R01HL119439-01A1 (to J.D.D.) from the NIH. Proteomics analysis was performed at the Northwestern Proteomics Core Facility, supported by the grants NCI CCSG P30 CA060553 and P41 GM108569 from the NIH.

Keywords

copA
nascent peptide
pseudoknot
recoding
ribosome profiling
slippery sequence
translation regulation

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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10.1016/j.molcel.2016.12.008

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title = "Programmed Ribosomal Frameshifting Generates a Copper Transporter and a Copper Chaperone from the Same Gene",

abstract = "Metal efflux pumps maintain ion homeostasis in the cell. The functions of the transporters are often supported by chaperone proteins, which scavenge the metal ions from the cytoplasm. Although the copper ion transporter CopA has been known in Escherichia coli, no gene for its chaperone had been identified. We show that the CopA chaperone is expressed in E. coli from the same gene that encodes the transporter. Some ribosomes translating copA undergo programmed frameshifting, terminate translation in the −1 frame, and generate the 70 aa-long polypeptide CopA(Z), which helps cells survive toxic copper concentrations. The high efficiency of frameshifting is achieved by the combined stimulatory action of a “slippery” sequence, an mRNA pseudoknot, and the CopA nascent chain. Similar mRNA elements are not only found in the copA genes of other bacteria but are also present in ATP7B, the human homolog of copA, and direct ribosomal frameshifting in vivo.",

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author = "Sezen Meydan and Dorota Klepacki and Subbulakshmi Karthikeyan and T{\~o}nu Margus and Paul Thomas and Jones, {John E.} and Yousuf Khan and Joseph Briggs and Dinman, {Jonathan D.} and Nora V{\'a}zquez-Laslop and Mankin, {Alexander S.}",

note = "Funding Information: We thank Pavel Baranov and John Atkins for helpful advice; Kurt Fredrick and Hyunwoo Lee for providing the pEK4 and pKOV plasmids, respectively; Andrew Jin for help with some experiments; and Dimple Modi, Nikolay Aleksashin, and Hao Lei for advice with some experimental procedures. This work was supported by the grants MCB 1244455 and MCB 1615851 (both to A.S.M. and N.V.-L.) from the National Science Foundation and R01 GM117177 and R01HL119439-01A1 (to J.D.D.) from the NIH. Proteomics analysis was performed at the Northwestern Proteomics Core Facility, supported by the grants NCI CCSG P30 CA060553 and P41 GM108569 from the NIH. Publisher Copyright: {\textcopyright} 2017 Elsevier Inc.",

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AU - Meydan, Sezen

AU - Klepacki, Dorota

AU - Karthikeyan, Subbulakshmi

AU - Margus, Tõnu

AU - Thomas, Paul

AU - Jones, John E.

AU - Khan, Yousuf

AU - Briggs, Joseph

AU - Dinman, Jonathan D.

AU - Vázquez-Laslop, Nora

AU - Mankin, Alexander S.

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PY - 2017/1/19

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N2 - Metal efflux pumps maintain ion homeostasis in the cell. The functions of the transporters are often supported by chaperone proteins, which scavenge the metal ions from the cytoplasm. Although the copper ion transporter CopA has been known in Escherichia coli, no gene for its chaperone had been identified. We show that the CopA chaperone is expressed in E. coli from the same gene that encodes the transporter. Some ribosomes translating copA undergo programmed frameshifting, terminate translation in the −1 frame, and generate the 70 aa-long polypeptide CopA(Z), which helps cells survive toxic copper concentrations. The high efficiency of frameshifting is achieved by the combined stimulatory action of a “slippery” sequence, an mRNA pseudoknot, and the CopA nascent chain. Similar mRNA elements are not only found in the copA genes of other bacteria but are also present in ATP7B, the human homolog of copA, and direct ribosomal frameshifting in vivo.

AB - Metal efflux pumps maintain ion homeostasis in the cell. The functions of the transporters are often supported by chaperone proteins, which scavenge the metal ions from the cytoplasm. Although the copper ion transporter CopA has been known in Escherichia coli, no gene for its chaperone had been identified. We show that the CopA chaperone is expressed in E. coli from the same gene that encodes the transporter. Some ribosomes translating copA undergo programmed frameshifting, terminate translation in the −1 frame, and generate the 70 aa-long polypeptide CopA(Z), which helps cells survive toxic copper concentrations. The high efficiency of frameshifting is achieved by the combined stimulatory action of a “slippery” sequence, an mRNA pseudoknot, and the CopA nascent chain. Similar mRNA elements are not only found in the copA genes of other bacteria but are also present in ATP7B, the human homolog of copA, and direct ribosomal frameshifting in vivo.

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KW - translation regulation

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Programmed Ribosomal Frameshifting Generates a Copper Transporter and a Copper Chaperone from the Same Gene

Abstract

Funding

Keywords

ASJC Scopus subject areas

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