Protein body formation in leaves of Nicotiana benthamiana: A concentration-dependent mechanism influenced by the presence of fusion tags

Reza Saberianfar, Jussi J. Joensuu, Andrew J. Conley, Rima Menassa (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

15 Citations (Scopus)

Abstract

Protein bodies (PBs) are endoplasmic reticulum (ER) derived organelles originally found in seeds whose function is to accumulate seed storage proteins. It has been shown that PB formation is not limited to seeds and green fluorescent protein (GFP) fused to either elastin-like polypeptide (ELP) or hydrophobin (HFBI) fusion tags induce the formation of PBs in leaves of N. benthamiana. In this study, we compared the ELP- and HFBI-induced PBs and showed that ELP-induced PBs are larger than HFBI-induced PBs. The size of ELP- and HFBI-induced PBs increased over time along with the accumulation levels of their fused protein. Our results show that PB formation is a concentration-dependent mechanism in which proteins accumulating at levels higher than 0.2% of total soluble protein are capable of inducing PBs in vivo. Our results show that the presence of fusion tags is not necessary for the formation of PBs, but affects the distribution pattern and size of PBs. This was confirmed by PBs induced by fluorescent proteins as well as fungal xylanases. We noticed that in the process of PB formation, secretory and ER-resident molecules are passively sequestered into the lumen of PBs. We propose to use this property of PBs as a tool to increase the accumulation levels of erythropoietin and human interleukin-10 by co-expression with PB-inducing proteins.
Original languageEnglish
Pages (from-to)927-937
JournalPlant Biotechnology Journal
Volume13
Issue number7
DOIs
Publication statusPublished - 2015
MoE publication typeA1 Journal article-refereed

Fingerprint

protein bodies
Nicotiana benthamiana
Tobacco
leaves
Proteins
elastin
Elastin
polypeptides
endoplasmic reticulum
Peptides
Body Size
proteins
Endoplasmic Reticulum
Seeds
erythropoietin
seed storage proteins
Seed Storage Proteins
xylanases
interleukin-10
seeds

Keywords

  • Nicotiana benthamiana
  • elastin-like polypeptides
  • hydrophobin
  • PB
  • protein body formation
  • protein body
  • transient expression

Cite this

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title = "Protein body formation in leaves of Nicotiana benthamiana: A concentration-dependent mechanism influenced by the presence of fusion tags",
abstract = "Protein bodies (PBs) are endoplasmic reticulum (ER) derived organelles originally found in seeds whose function is to accumulate seed storage proteins. It has been shown that PB formation is not limited to seeds and green fluorescent protein (GFP) fused to either elastin-like polypeptide (ELP) or hydrophobin (HFBI) fusion tags induce the formation of PBs in leaves of N. benthamiana. In this study, we compared the ELP- and HFBI-induced PBs and showed that ELP-induced PBs are larger than HFBI-induced PBs. The size of ELP- and HFBI-induced PBs increased over time along with the accumulation levels of their fused protein. Our results show that PB formation is a concentration-dependent mechanism in which proteins accumulating at levels higher than 0.2{\%} of total soluble protein are capable of inducing PBs in vivo. Our results show that the presence of fusion tags is not necessary for the formation of PBs, but affects the distribution pattern and size of PBs. This was confirmed by PBs induced by fluorescent proteins as well as fungal xylanases. We noticed that in the process of PB formation, secretory and ER-resident molecules are passively sequestered into the lumen of PBs. We propose to use this property of PBs as a tool to increase the accumulation levels of erythropoietin and human interleukin-10 by co-expression with PB-inducing proteins.",
keywords = "Nicotiana benthamiana, elastin-like polypeptides, hydrophobin, PB, protein body formation, protein body, transient expression",
author = "Reza Saberianfar and Joensuu, {Jussi J.} and Conley, {Andrew J.} and Rima Menassa",
year = "2015",
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journal = "Plant Biotechnology Journal",
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Protein body formation in leaves of Nicotiana benthamiana: A concentration-dependent mechanism influenced by the presence of fusion tags. / Saberianfar, Reza; Joensuu, Jussi J.; Conley, Andrew J.; Menassa, Rima (Corresponding Author).

In: Plant Biotechnology Journal, Vol. 13, No. 7, 2015, p. 927-937.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Protein body formation in leaves of Nicotiana benthamiana: A concentration-dependent mechanism influenced by the presence of fusion tags

AU - Saberianfar, Reza

AU - Joensuu, Jussi J.

AU - Conley, Andrew J.

AU - Menassa, Rima

PY - 2015

Y1 - 2015

N2 - Protein bodies (PBs) are endoplasmic reticulum (ER) derived organelles originally found in seeds whose function is to accumulate seed storage proteins. It has been shown that PB formation is not limited to seeds and green fluorescent protein (GFP) fused to either elastin-like polypeptide (ELP) or hydrophobin (HFBI) fusion tags induce the formation of PBs in leaves of N. benthamiana. In this study, we compared the ELP- and HFBI-induced PBs and showed that ELP-induced PBs are larger than HFBI-induced PBs. The size of ELP- and HFBI-induced PBs increased over time along with the accumulation levels of their fused protein. Our results show that PB formation is a concentration-dependent mechanism in which proteins accumulating at levels higher than 0.2% of total soluble protein are capable of inducing PBs in vivo. Our results show that the presence of fusion tags is not necessary for the formation of PBs, but affects the distribution pattern and size of PBs. This was confirmed by PBs induced by fluorescent proteins as well as fungal xylanases. We noticed that in the process of PB formation, secretory and ER-resident molecules are passively sequestered into the lumen of PBs. We propose to use this property of PBs as a tool to increase the accumulation levels of erythropoietin and human interleukin-10 by co-expression with PB-inducing proteins.

AB - Protein bodies (PBs) are endoplasmic reticulum (ER) derived organelles originally found in seeds whose function is to accumulate seed storage proteins. It has been shown that PB formation is not limited to seeds and green fluorescent protein (GFP) fused to either elastin-like polypeptide (ELP) or hydrophobin (HFBI) fusion tags induce the formation of PBs in leaves of N. benthamiana. In this study, we compared the ELP- and HFBI-induced PBs and showed that ELP-induced PBs are larger than HFBI-induced PBs. The size of ELP- and HFBI-induced PBs increased over time along with the accumulation levels of their fused protein. Our results show that PB formation is a concentration-dependent mechanism in which proteins accumulating at levels higher than 0.2% of total soluble protein are capable of inducing PBs in vivo. Our results show that the presence of fusion tags is not necessary for the formation of PBs, but affects the distribution pattern and size of PBs. This was confirmed by PBs induced by fluorescent proteins as well as fungal xylanases. We noticed that in the process of PB formation, secretory and ER-resident molecules are passively sequestered into the lumen of PBs. We propose to use this property of PBs as a tool to increase the accumulation levels of erythropoietin and human interleukin-10 by co-expression with PB-inducing proteins.

KW - Nicotiana benthamiana

KW - elastin-like polypeptides

KW - hydrophobin

KW - PB

KW - protein body formation

KW - protein body

KW - transient expression

U2 - 10.1111/pbi.12329

DO - 10.1111/pbi.12329

M3 - Article

VL - 13

SP - 927

EP - 937

JO - Plant Biotechnology Journal

JF - Plant Biotechnology Journal

SN - 1467-7644

IS - 7

ER -