Functional significance of AtHMA4 C-terminal domain in planta

Rebecca F Mills, Billy Valdes, Michael Duke, Kerry A Peaston, Brett Lahner, David E Salt, Lorraine E Williams

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Abstract

Background: Enhancing the upward translocation of heavy metals such as Zn from root to shoot through genetic engineering has potential for biofortification and phytoremediation. This study examined the contribution of the heavy metal-transporting ATPase, AtHMA4, to the shoot ionomic profile of soil-grown plants, and investigated the importance of the C-terminal domain in the functioning of this transporter.

Principal Findings: The Arabidopsis hma2 hma4 mutant has a stunted phenotype and a distinctive ionomic profile, with low shoot levels of Zn, Cd, Co, K and Rb, and high shoot Cu. Expression of AtHMA4 (AtHMA4-FL) under the CaMV-35S promoter partially rescued the stunted phenotype of hma2 hma4; rosette diameter returned to wild-type levels in the majority of lines and bolts were also produced, although the average bolt height was not restored completely. AtHMA4-FL expression rescued Co, K, Rb and Cu to wild-type levels, and partially returned Cd and Zn levels (83% and 28% of wild type respectively). In contrast, expression of AtHMA4-trunc (without the C-terminal region) in hma2 hma4 only partially restored the rosette diameter in two of five lines and bolt production was not rescued. There was no significant effect on the shoot ionomic profile, apart from Cd, which was increased to 41% of wild-type levels. When the AtHMA4 C-terminal domain (AtHMA4-C-term) was expressed in hma2 hma4 it had no marked effect. When expressed in yeast, AtHMA4-C-term and AtHMA4-trunc conferred greater Cd and Zn tolerance than AtHMA4-FL.

Conclusion: The ionome of the hma2 hma4 mutant differs markedly from wt plants. The functional relevance of domains of AtHMA4 in planta can be explored by complementing this mutant. AtHMA4-FL is more effective in restoring shoot metal accumulation in this mutant than a C-terminally truncated version of the pump, indicating that the C-terminal domain is important in the functioning of AtHMA4 in planta.

Original languageEnglish
Article numbere13388
Number of pages10
JournalPloS ONE
Volume5
Issue number10
DOIs
Publication statusPublished - 20 Oct 2010

Keywords

  • metal-binding domain
  • arabidopsis-thaliana
  • thlaspi-caerulescens
  • transporting atpase
  • zinc homeostasis
  • P-1B-type aptase
  • cadmium
  • expression
  • hyperaccumulation
  • biofortification

Cite this

Mills, R. F., Valdes, B., Duke, M., Peaston, K. A., Lahner, B., Salt, D. E., & Williams, L. E. (2010). Functional significance of AtHMA4 C-terminal domain in planta. PloS ONE, 5(10), [e13388]. https://doi.org/10.1371/journal.pone.0013388

Functional significance of AtHMA4 C-terminal domain in planta. / Mills, Rebecca F; Valdes, Billy; Duke, Michael; Peaston, Kerry A; Lahner, Brett; Salt, David E; Williams, Lorraine E.

In: PloS ONE, Vol. 5, No. 10, e13388, 20.10.2010.

Research output: Contribution to journalArticle

Mills, RF, Valdes, B, Duke, M, Peaston, KA, Lahner, B, Salt, DE & Williams, LE 2010, 'Functional significance of AtHMA4 C-terminal domain in planta' PloS ONE, vol. 5, no. 10, e13388. https://doi.org/10.1371/journal.pone.0013388
Mills RF, Valdes B, Duke M, Peaston KA, Lahner B, Salt DE et al. Functional significance of AtHMA4 C-terminal domain in planta. PloS ONE. 2010 Oct 20;5(10). e13388. https://doi.org/10.1371/journal.pone.0013388
Mills, Rebecca F ; Valdes, Billy ; Duke, Michael ; Peaston, Kerry A ; Lahner, Brett ; Salt, David E ; Williams, Lorraine E. / Functional significance of AtHMA4 C-terminal domain in planta. In: PloS ONE. 2010 ; Vol. 5, No. 10.
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abstract = "Background: Enhancing the upward translocation of heavy metals such as Zn from root to shoot through genetic engineering has potential for biofortification and phytoremediation. This study examined the contribution of the heavy metal-transporting ATPase, AtHMA4, to the shoot ionomic profile of soil-grown plants, and investigated the importance of the C-terminal domain in the functioning of this transporter.Principal Findings: The Arabidopsis hma2 hma4 mutant has a stunted phenotype and a distinctive ionomic profile, with low shoot levels of Zn, Cd, Co, K and Rb, and high shoot Cu. Expression of AtHMA4 (AtHMA4-FL) under the CaMV-35S promoter partially rescued the stunted phenotype of hma2 hma4; rosette diameter returned to wild-type levels in the majority of lines and bolts were also produced, although the average bolt height was not restored completely. AtHMA4-FL expression rescued Co, K, Rb and Cu to wild-type levels, and partially returned Cd and Zn levels (83{\%} and 28{\%} of wild type respectively). In contrast, expression of AtHMA4-trunc (without the C-terminal region) in hma2 hma4 only partially restored the rosette diameter in two of five lines and bolt production was not rescued. There was no significant effect on the shoot ionomic profile, apart from Cd, which was increased to 41{\%} of wild-type levels. When the AtHMA4 C-terminal domain (AtHMA4-C-term) was expressed in hma2 hma4 it had no marked effect. When expressed in yeast, AtHMA4-C-term and AtHMA4-trunc conferred greater Cd and Zn tolerance than AtHMA4-FL.Conclusion: The ionome of the hma2 hma4 mutant differs markedly from wt plants. The functional relevance of domains of AtHMA4 in planta can be explored by complementing this mutant. AtHMA4-FL is more effective in restoring shoot metal accumulation in this mutant than a C-terminally truncated version of the pump, indicating that the C-terminal domain is important in the functioning of AtHMA4 in planta.",
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T1 - Functional significance of AtHMA4 C-terminal domain in planta

AU - Mills, Rebecca F

AU - Valdes, Billy

AU - Duke, Michael

AU - Peaston, Kerry A

AU - Lahner, Brett

AU - Salt, David E

AU - Williams, Lorraine E

PY - 2010/10/20

Y1 - 2010/10/20

N2 - Background: Enhancing the upward translocation of heavy metals such as Zn from root to shoot through genetic engineering has potential for biofortification and phytoremediation. This study examined the contribution of the heavy metal-transporting ATPase, AtHMA4, to the shoot ionomic profile of soil-grown plants, and investigated the importance of the C-terminal domain in the functioning of this transporter.Principal Findings: The Arabidopsis hma2 hma4 mutant has a stunted phenotype and a distinctive ionomic profile, with low shoot levels of Zn, Cd, Co, K and Rb, and high shoot Cu. Expression of AtHMA4 (AtHMA4-FL) under the CaMV-35S promoter partially rescued the stunted phenotype of hma2 hma4; rosette diameter returned to wild-type levels in the majority of lines and bolts were also produced, although the average bolt height was not restored completely. AtHMA4-FL expression rescued Co, K, Rb and Cu to wild-type levels, and partially returned Cd and Zn levels (83% and 28% of wild type respectively). In contrast, expression of AtHMA4-trunc (without the C-terminal region) in hma2 hma4 only partially restored the rosette diameter in two of five lines and bolt production was not rescued. There was no significant effect on the shoot ionomic profile, apart from Cd, which was increased to 41% of wild-type levels. When the AtHMA4 C-terminal domain (AtHMA4-C-term) was expressed in hma2 hma4 it had no marked effect. When expressed in yeast, AtHMA4-C-term and AtHMA4-trunc conferred greater Cd and Zn tolerance than AtHMA4-FL.Conclusion: The ionome of the hma2 hma4 mutant differs markedly from wt plants. The functional relevance of domains of AtHMA4 in planta can be explored by complementing this mutant. AtHMA4-FL is more effective in restoring shoot metal accumulation in this mutant than a C-terminally truncated version of the pump, indicating that the C-terminal domain is important in the functioning of AtHMA4 in planta.

AB - Background: Enhancing the upward translocation of heavy metals such as Zn from root to shoot through genetic engineering has potential for biofortification and phytoremediation. This study examined the contribution of the heavy metal-transporting ATPase, AtHMA4, to the shoot ionomic profile of soil-grown plants, and investigated the importance of the C-terminal domain in the functioning of this transporter.Principal Findings: The Arabidopsis hma2 hma4 mutant has a stunted phenotype and a distinctive ionomic profile, with low shoot levels of Zn, Cd, Co, K and Rb, and high shoot Cu. Expression of AtHMA4 (AtHMA4-FL) under the CaMV-35S promoter partially rescued the stunted phenotype of hma2 hma4; rosette diameter returned to wild-type levels in the majority of lines and bolts were also produced, although the average bolt height was not restored completely. AtHMA4-FL expression rescued Co, K, Rb and Cu to wild-type levels, and partially returned Cd and Zn levels (83% and 28% of wild type respectively). In contrast, expression of AtHMA4-trunc (without the C-terminal region) in hma2 hma4 only partially restored the rosette diameter in two of five lines and bolt production was not rescued. There was no significant effect on the shoot ionomic profile, apart from Cd, which was increased to 41% of wild-type levels. When the AtHMA4 C-terminal domain (AtHMA4-C-term) was expressed in hma2 hma4 it had no marked effect. When expressed in yeast, AtHMA4-C-term and AtHMA4-trunc conferred greater Cd and Zn tolerance than AtHMA4-FL.Conclusion: The ionome of the hma2 hma4 mutant differs markedly from wt plants. The functional relevance of domains of AtHMA4 in planta can be explored by complementing this mutant. AtHMA4-FL is more effective in restoring shoot metal accumulation in this mutant than a C-terminally truncated version of the pump, indicating that the C-terminal domain is important in the functioning of AtHMA4 in planta.

KW - metal-binding domain

KW - arabidopsis-thaliana

KW - thlaspi-caerulescens

KW - transporting atpase

KW - zinc homeostasis

KW - P-1B-type aptase

KW - cadmium

KW - expression

KW - hyperaccumulation

KW - biofortification

U2 - 10.1371/journal.pone.0013388

DO - 10.1371/journal.pone.0013388

M3 - Article

VL - 5

JO - PloS ONE

JF - PloS ONE

SN - 1932-6203

IS - 10

M1 - e13388

ER -