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Four signature motifs define the first class of structurally related large coiled-coil proteins in plants.

Gindullis F, Rose A, Patel S, Meier I - BMC Genomics (2002)

Bottom Line: Filament-like plant proteins (FPP) were identified by sequence similarity to a tomato cDNA that encodes a coiled-coil protein which interacts with the nuclear envelope-associated protein, MAF1.The tomato homolog of the FPP family binds in a yeast two-hybrid assay to a nuclear envelope-associated protein.Because the full Arabidopsis genome does not appear to contain genes for lamins, it is of interest to investigate other long coiled-coil proteins, which might functionally replace lamins in the plant kingdom.

View Article: PubMed Central - HTML - PubMed

Affiliation: CellTec Biotechnologie GmbH, Frohmestrasse 110, D-22459 Hamburg, Germany. frank.gindullis@celltec.de

ABSTRACT

Background: Animal and yeast proteins containing long coiled-coil domains are involved in attaching other proteins to the large, solid-state components of the cell. One subgroup of long coiled-coil proteins are the nuclear lamins, which are involved in attaching chromatin to the nuclear envelope and have recently been implicated in inherited human diseases. In contrast to other eukaryotes, long coiled-coil proteins have been barely investigated in plants.

Results: We have searched the completed Arabidopsis genome and have identified a family of structurally related long coiled-coil proteins. Filament-like plant proteins (FPP) were identified by sequence similarity to a tomato cDNA that encodes a coiled-coil protein which interacts with the nuclear envelope-associated protein, MAF1. The FPP family is defined by four novel unique sequence motifs and by two clusters of long coiled-coil domains separated by a non-coiled-coil linker. All family members are expressed in a variety of Arabidopsis tissues. A homolog sharing the structural features was identified in the monocot rice, indicating conservation among angiosperms.

Conclusion: Except for myosins, this is the first characterization of a family of long coiled-coil proteins in plants. The tomato homolog of the FPP family binds in a yeast two-hybrid assay to a nuclear envelope-associated protein. This might suggest that FPP family members function in nuclear envelope biology. Because the full Arabidopsis genome does not appear to contain genes for lamins, it is of interest to investigate other long coiled-coil proteins, which might functionally replace lamins in the plant kingdom.

No MeSH data available.


Yeast two-hybrid interaction of LeFPP with MAF1. (A)Yeast colonies co-expressing bait and prey plasmids were streaked on dropout plates without leucine and tryptophane (LT) or dropout plates without leucine, tryptophane, and histidine (LTH). (a) pBD-LeMAF1/pAD-LeFPP; (b) pBD-AtMAF1/pAD-LeFPP; (c) pBD-AtRanGAP1/pAD-LeFPP; (d) pBD-GAL4/pAD-LeFPP. (B) Quantification of β-Galactosidase activity in yeast strains harboring combinations of bait and prey plasmids. (1) p53/pSV40 (positive control); (2) pLaminC/pSV40 (negative control); (3) pBD-LeMAF1/pAD-LeFPP; (4) pBD-GAL4/pAD-LeFPP. Units, relative β-Galactosidase units (see Methods). (C) Sequence alignment between LeMAF1, AtMAF1, and the N-terminal domain of AtRanGAP1. Identical amino acids are shaded in black.
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Figure 1: Yeast two-hybrid interaction of LeFPP with MAF1. (A)Yeast colonies co-expressing bait and prey plasmids were streaked on dropout plates without leucine and tryptophane (LT) or dropout plates without leucine, tryptophane, and histidine (LTH). (a) pBD-LeMAF1/pAD-LeFPP; (b) pBD-AtMAF1/pAD-LeFPP; (c) pBD-AtRanGAP1/pAD-LeFPP; (d) pBD-GAL4/pAD-LeFPP. (B) Quantification of β-Galactosidase activity in yeast strains harboring combinations of bait and prey plasmids. (1) p53/pSV40 (positive control); (2) pLaminC/pSV40 (negative control); (3) pBD-LeMAF1/pAD-LeFPP; (4) pBD-GAL4/pAD-LeFPP. Units, relative β-Galactosidase units (see Methods). (C) Sequence alignment between LeMAF1, AtMAF1, and the N-terminal domain of AtRanGAP1. Identical amino acids are shaded in black.

Mentions: A yeast two-hybrid screen was performed to identify interaction partners of the tomato nuclear envelope-associated protein LeMAF1 [36]. A prey plasmid (pAD-LeFPP) was identified that led to activation of the two reporter genes HIS and LacZ after retransformation with the pBD-LeMAF1 bait. We tested the specificity of this interaction by co-transforming pAD-LeFPP with two additional bait plasmids, pBD-AtMAF1 and pBD-AtRanGAP1. AtMAF1 is one of three Arabidopsis homologs of LeMAF1 (Patel and Meier, unpublished results), and has 45% amino acid identity with LeMAF1. AtRanGAP1 is one of the two Arabidopsis Ran GTPase activating proteins. Its N-terminus has 28% amino acid identity with LeMAF1 and 30% with AtMAF1 (Fig. 1C; [33,37]). Fig. 1A shows that pAD-LeFPP leads to activation of the HIS gene in combination with pBD-LeMAF1 (sector a) and with pBD-AtMAF1 (sector b), but not with pBDAtRanGAP1 (sector c) or the binding domain vector pBD-GAL4 alone (sector d). This demonstrates that the observed interaction is specific for MAF1-like proteins.


Four signature motifs define the first class of structurally related large coiled-coil proteins in plants.

Gindullis F, Rose A, Patel S, Meier I - BMC Genomics (2002)

Yeast two-hybrid interaction of LeFPP with MAF1. (A)Yeast colonies co-expressing bait and prey plasmids were streaked on dropout plates without leucine and tryptophane (LT) or dropout plates without leucine, tryptophane, and histidine (LTH). (a) pBD-LeMAF1/pAD-LeFPP; (b) pBD-AtMAF1/pAD-LeFPP; (c) pBD-AtRanGAP1/pAD-LeFPP; (d) pBD-GAL4/pAD-LeFPP. (B) Quantification of β-Galactosidase activity in yeast strains harboring combinations of bait and prey plasmids. (1) p53/pSV40 (positive control); (2) pLaminC/pSV40 (negative control); (3) pBD-LeMAF1/pAD-LeFPP; (4) pBD-GAL4/pAD-LeFPP. Units, relative β-Galactosidase units (see Methods). (C) Sequence alignment between LeMAF1, AtMAF1, and the N-terminal domain of AtRanGAP1. Identical amino acids are shaded in black.
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Related In: Results  -  Collection

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Figure 1: Yeast two-hybrid interaction of LeFPP with MAF1. (A)Yeast colonies co-expressing bait and prey plasmids were streaked on dropout plates without leucine and tryptophane (LT) or dropout plates without leucine, tryptophane, and histidine (LTH). (a) pBD-LeMAF1/pAD-LeFPP; (b) pBD-AtMAF1/pAD-LeFPP; (c) pBD-AtRanGAP1/pAD-LeFPP; (d) pBD-GAL4/pAD-LeFPP. (B) Quantification of β-Galactosidase activity in yeast strains harboring combinations of bait and prey plasmids. (1) p53/pSV40 (positive control); (2) pLaminC/pSV40 (negative control); (3) pBD-LeMAF1/pAD-LeFPP; (4) pBD-GAL4/pAD-LeFPP. Units, relative β-Galactosidase units (see Methods). (C) Sequence alignment between LeMAF1, AtMAF1, and the N-terminal domain of AtRanGAP1. Identical amino acids are shaded in black.
Mentions: A yeast two-hybrid screen was performed to identify interaction partners of the tomato nuclear envelope-associated protein LeMAF1 [36]. A prey plasmid (pAD-LeFPP) was identified that led to activation of the two reporter genes HIS and LacZ after retransformation with the pBD-LeMAF1 bait. We tested the specificity of this interaction by co-transforming pAD-LeFPP with two additional bait plasmids, pBD-AtMAF1 and pBD-AtRanGAP1. AtMAF1 is one of three Arabidopsis homologs of LeMAF1 (Patel and Meier, unpublished results), and has 45% amino acid identity with LeMAF1. AtRanGAP1 is one of the two Arabidopsis Ran GTPase activating proteins. Its N-terminus has 28% amino acid identity with LeMAF1 and 30% with AtMAF1 (Fig. 1C; [33,37]). Fig. 1A shows that pAD-LeFPP leads to activation of the HIS gene in combination with pBD-LeMAF1 (sector a) and with pBD-AtMAF1 (sector b), but not with pBDAtRanGAP1 (sector c) or the binding domain vector pBD-GAL4 alone (sector d). This demonstrates that the observed interaction is specific for MAF1-like proteins.

Bottom Line: Filament-like plant proteins (FPP) were identified by sequence similarity to a tomato cDNA that encodes a coiled-coil protein which interacts with the nuclear envelope-associated protein, MAF1.The tomato homolog of the FPP family binds in a yeast two-hybrid assay to a nuclear envelope-associated protein.Because the full Arabidopsis genome does not appear to contain genes for lamins, it is of interest to investigate other long coiled-coil proteins, which might functionally replace lamins in the plant kingdom.

View Article: PubMed Central - HTML - PubMed

Affiliation: CellTec Biotechnologie GmbH, Frohmestrasse 110, D-22459 Hamburg, Germany. frank.gindullis@celltec.de

ABSTRACT

Background: Animal and yeast proteins containing long coiled-coil domains are involved in attaching other proteins to the large, solid-state components of the cell. One subgroup of long coiled-coil proteins are the nuclear lamins, which are involved in attaching chromatin to the nuclear envelope and have recently been implicated in inherited human diseases. In contrast to other eukaryotes, long coiled-coil proteins have been barely investigated in plants.

Results: We have searched the completed Arabidopsis genome and have identified a family of structurally related long coiled-coil proteins. Filament-like plant proteins (FPP) were identified by sequence similarity to a tomato cDNA that encodes a coiled-coil protein which interacts with the nuclear envelope-associated protein, MAF1. The FPP family is defined by four novel unique sequence motifs and by two clusters of long coiled-coil domains separated by a non-coiled-coil linker. All family members are expressed in a variety of Arabidopsis tissues. A homolog sharing the structural features was identified in the monocot rice, indicating conservation among angiosperms.

Conclusion: Except for myosins, this is the first characterization of a family of long coiled-coil proteins in plants. The tomato homolog of the FPP family binds in a yeast two-hybrid assay to a nuclear envelope-associated protein. This might suggest that FPP family members function in nuclear envelope biology. Because the full Arabidopsis genome does not appear to contain genes for lamins, it is of interest to investigate other long coiled-coil proteins, which might functionally replace lamins in the plant kingdom.

No MeSH data available.