The molecular characterisation of type IIA calcium-atpases in arabidopsis thaliana
The molecular characterisation of type IIA calcium-atpases in arabidopsis thaliana
Two type IIA (SERCA-type) Ca2+-ATPases were identified in Arabidopsis thaliana ecotype Landsberg erecta, in addition to a previously identified pump, AtECA1. The identification of two additional clones was achieved using a PCR-approach. Two full-length of cDNA clones AtECA2 and AtECA3 (Arabidopsis thaliana ER-type Ca2+-ATPase 2 and 3) were amplified and sequenced. AtECA2 encodes a protein of 116 kDa and AtECA3 encodes a smaller protein of 109 kDa. Detailed sequence analysis compared these clones with AtECA1 of Arabidopsis, and other eukaryotic Ca2+-ATPases. The three Arabidopsis Ca2+ pumps are members of the type IIA family of P-type ATPases. AtECA1 and AtECA2 are closely related genes which have high sequence similarity to other plant type IIA Ca2+-ATPases. However AtECA3 shows greater similarity to animal SERCA pumps than other plant homologues. Phylogenetic analysis of Ca2+-ATPases from various species also suggest that the type IIA group can be further divided into sub-groups.
Southern analysis suggests that each of the Arabidopsis type IIA Ca2+-ATPase genes are present as a single copy and indicates that a small family of moderately related type IIA genes are present in this plant. Expression analysis using RT-PCR demonstrated that the three type IIA pumps have distinct organ-specific expression patterns. Both AtECA1 and AtECA3 are present in all major organs, while AtECA2 is strongly expressed in stem tissue but is present at very low levels in root.
AtECA1 and SERCA1, a fast-twitch muscle Ca2+-ATPase from rabbit, were expressed in a yeast strain which is deficient in two endogenous Ca2+-ATPases PMR1 and PMC1. Both Ca2+-ATPases restored the growth of the mutant yeast strain, indicating that they can transport calcium. Three type IIA Ca2+-ATPases are expressed in Arabidopsis. The identification of these clones will provide further molecular tools with which to help to understand the regulation of calcium signalling events in this plant.
University of Southampton
1999
Pittman, Jon Kevin
(1999)
The molecular characterisation of type IIA calcium-atpases in arabidopsis thaliana.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
Two type IIA (SERCA-type) Ca2+-ATPases were identified in Arabidopsis thaliana ecotype Landsberg erecta, in addition to a previously identified pump, AtECA1. The identification of two additional clones was achieved using a PCR-approach. Two full-length of cDNA clones AtECA2 and AtECA3 (Arabidopsis thaliana ER-type Ca2+-ATPase 2 and 3) were amplified and sequenced. AtECA2 encodes a protein of 116 kDa and AtECA3 encodes a smaller protein of 109 kDa. Detailed sequence analysis compared these clones with AtECA1 of Arabidopsis, and other eukaryotic Ca2+-ATPases. The three Arabidopsis Ca2+ pumps are members of the type IIA family of P-type ATPases. AtECA1 and AtECA2 are closely related genes which have high sequence similarity to other plant type IIA Ca2+-ATPases. However AtECA3 shows greater similarity to animal SERCA pumps than other plant homologues. Phylogenetic analysis of Ca2+-ATPases from various species also suggest that the type IIA group can be further divided into sub-groups.
Southern analysis suggests that each of the Arabidopsis type IIA Ca2+-ATPase genes are present as a single copy and indicates that a small family of moderately related type IIA genes are present in this plant. Expression analysis using RT-PCR demonstrated that the three type IIA pumps have distinct organ-specific expression patterns. Both AtECA1 and AtECA3 are present in all major organs, while AtECA2 is strongly expressed in stem tissue but is present at very low levels in root.
AtECA1 and SERCA1, a fast-twitch muscle Ca2+-ATPase from rabbit, were expressed in a yeast strain which is deficient in two endogenous Ca2+-ATPases PMR1 and PMC1. Both Ca2+-ATPases restored the growth of the mutant yeast strain, indicating that they can transport calcium. Three type IIA Ca2+-ATPases are expressed in Arabidopsis. The identification of these clones will provide further molecular tools with which to help to understand the regulation of calcium signalling events in this plant.
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Published date: 1999
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Local EPrints ID: 464005
URI: http://eprints.soton.ac.uk/id/eprint/464005
PURE UUID: 1f7f5d1c-e6d7-4ee8-87b2-409a3e24b240
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Date deposited: 04 Jul 2022 21:00
Last modified: 04 Jul 2022 21:00
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Author:
Jon Kevin Pittman
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