The C-terminal tetrapeptide of phaseolin is sufficient to target green fluorescent protein to the vacuole

Frigerio, Lorenzo; Foresti, Ombretta; Felipe, Doramys Hernández; Neuhaus, Jean‐Marc; Vitale, Alessandro

doi:10.1078/0176-1617-00362

Cited by 42 publications

(54 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…uole (Frigerio et al, 2001a). After 1 h of pulse labeling, neither of the two GFP constructs could be coselected using anti-BiP antiserum ( Figure 3, lanes 5 to 8), indicating that permanence in the ER does not result directly in the interaction with BiP detected by our assay and that the addition of the vacuolar sorting signal of phaseolin does not stimulate interactions with BiP.…”

Section: The C-terminal Domain Of Phaseolin Stimulates the Interactiomentioning

confidence: 92%

“…GFP is a small soluble protein that is secreted efficiently when introduced into the plant ER via a signal peptide (Batoko et al, 2000;Frigerio et al, 2001a). A number of C-terminal additions have been performed on ER-introduced GFP without affecting its fluorescence, and therefore its folding, in plant cells (Haseloff et al, 1997;Di Sansebastiano et al, 1998;Benghezal et al, 2000;Frigerio et al, 2001a).…”

Section: The C-terminal Domain Of Phaseolin Stimulates the Interactiomentioning

confidence: 99%

“…sGFPAFVY has been described (Frigerio et al, 2001a). sGFPHDEL (plasmid pBIN-mGFP5-ER; a gift from Jim Haseloff, University of Cambridge, UK) contains the GFP5 coding sequence fused to the Arabidopsis thaliana basic chitinase signal peptide and the C-terminal endoplasmic reticulum (ER) retrieval signal HDEL.…”

Section: Recombinant Dnamentioning

confidence: 99%

“…tumefaciens LBA4404 containing sGFP418, sGFPHDEL, or AC1 was used to produce transgenic tobacco (Nicotiana tabacum cv Petit Havana SR1) plants as described by Pedrazzini et al (1997). The production of transgenic tobacco expressing sGFPAFVY has been described (Frigerio et al, 2001a).…”

Section: Production Of Transgenic Tobacco Plants and Transient Transfmentioning

confidence: 99%

See 3 more Smart Citations

A Phaseolin Domain Involved Directly in Trimer Assembly Is a Determinant for Binding by the Chaperone BiP

Foresti

Frigerio

Holkeri

et al. 2003

THE PLANT CELL ONLINE

View full text Add to dashboard Cite

The binding protein (BiP; a member of the heat-shock 70 family) is a major chaperone of the endoplasmic reticulum (ER). Interactions with BiP are believed to inhibit unproductive aggregation of newly synthesized secretory proteins during folding and assembly. In vitro, BiP has a preference for peptide sequences enriched in hydrophobic amino acids, which are expected to be exposed only in folding and assembly intermediates or in defective proteins. However, direct information regarding sequences recognized in vivo by BiP on real proteins is very limited. We have shown previously that newly synthesized monomers of the homotrimeric storage protein phaseolin associate with BiP and that phaseolin trimerization in the ER abolishes such interactions. Using different phaseolin constructs and green fluorescent protein (GFP) fusion proteins, we show here that one of the two ␣ -helical regions of polypeptide contact in phaseolin trimers (35 amino acids located close to the C terminus and containing three potential BiP binding sites) effectively promotes BiP association with phaseolin and with secretory GFP fusions expressed in transgenic tobacco or in transfected protoplasts. We also show that overexpressed BiP transiently sequesters phaseolin polypeptides. We conclude that one of the regions of monomer contact is a BiP binding determinant and suggest that during the synthesis of phaseolin, the association with BiP and trimer formation are competing events. Finally, we show that the other, internal region of contact between monomers is necessary for phaseolin assembly in vivo and contains one potential BiP binding site.

show abstract

Section: The C-terminal Domain Of Phaseolin Stimulates the Interactiomentioning

confidence: 92%

Section: The C-terminal Domain Of Phaseolin Stimulates the Interactiomentioning

confidence: 99%

Section: Recombinant Dnamentioning

confidence: 99%

Section: Production Of Transgenic Tobacco Plants and Transient Transfmentioning

confidence: 99%

See 2 more Smart Citations

A Phaseolin Domain Involved Directly in Trimer Assembly Is a Determinant for Binding by the Chaperone BiP

Foresti

Frigerio

Holkeri

et al. 2003

THE PLANT CELL ONLINE

View full text Add to dashboard Cite

show abstract

“…D418 is a mutated form of phaseolin characterized by the absence of the tetrapeptide AFVY, which is responsible for protein delivery to the vacuole in tobacco cells (Frigerio et al, 1998(Frigerio et al, , 2001a. In this case, the ManD418 fusion protein, instead of being secreted, was mainly localized in the ER as an intact polypeptide, but its processed fragments were detected in the vacuole, indicating that the MAN2B1 200-amino acid domain can at least partially redirect the secreted protein D418 to this organelle.…”

Section: Discussionmentioning

confidence: 99%

Traffic of Human α-Mannosidase in Plant Cells Suggests the Presence of a New Endoplasmic Reticulum-to-Vacuole Pathway without Involving the Golgi Complex

2013

View full text Add to dashboard Cite

The transport of secretory proteins from the endoplasmic reticulum to the vacuole requires sorting signals as well as specific transport mechanisms. This work is focused on the transport in transgenic tobacco (Nicotiana tabacum) plants of a human a-mannosidase, MAN2B1, which is a lysosomal enzyme involved in the turnover of N-linked glycoproteins and can be used in enzyme replacement therapy. Although ubiquitously expressed, a-mannosidases are targeted to lysosomes or vacuoles through different mechanisms according to the organisms in which these proteins are produced. In tobacco cells, MAN2B1 reaches the vacuole even in the absence of mannose-6-phosphate receptors, which are responsible for its transport in animal cells. We report that MAN2B1 is targeted to the vacuole without passing through the Golgi complex. In addition, a vacuolar targeting signal that is recognized in plant cells is located in the MAN2B1 amino-terminal region. Indeed, when this amino-terminal domain is removed, the protein is retained in the endoplasmic reticulum. Moreover, when this domain is added to a plantsecreted protein, the resulting fusion protein is partially redirected to the vacuole. These results strongly suggest the existence in plants of a new type of vacuolar traffic that can be used by leaf cells to transport vacuolar proteins.

show abstract

Plant Vacuoles: from Biogenesis to Function

Neuhaus¹,

Paris

2005

Plant Cell Monographs

View full text Add to dashboard Cite

The C-terminal tetrapeptide of phaseolin is sufficient to target green fluorescent protein to the vacuole

Cited by 42 publications

References 29 publications

A Phaseolin Domain Involved Directly in Trimer Assembly Is a Determinant for Binding by the Chaperone BiP

A Phaseolin Domain Involved Directly in Trimer Assembly Is a Determinant for Binding by the Chaperone BiP

Traffic of Human α-Mannosidase in Plant Cells Suggests the Presence of a New Endoplasmic Reticulum-to-Vacuole Pathway without Involving the Golgi Complex

Plant Vacuoles: from Biogenesis to Function

Contact Info

Product

Resources

About