1 Characterization and Functional Analysis of ABSCISIC ACID INSENSITIVE3-like Genes from Physcomitrella patens Heather H. Marella*, Yoichi Sakata 1 *, and Ralph S. Quatrano 2 Department of Biology, Washington University, 1 Brookings Dr., St. Louis, MO 63130, USA * These authors contributed equally to this work 2 Corresponding Author: Ralph Quatrano Address: Washington University, 1 Brookings Dr., Campus Box 1137, St. Louis, MO 63130 Email: [email protected]Telephone: 314-935-6850 Fax: 314-935-8137 Email of authors: Heather Marella- [email protected]Yoichi Sakata- [email protected]Running title: Physcomitrella patens ABI3 Key Words: ABI3, abscisic acid, Physcomitrella, ABI5, VP1, transcriptional regulation Accession Numbers: PpABI3A-AB233419, PpABI3B-AB233420, PpABI3C-AB245516 Word Count: Total-9875 Summary-250 Introduction-1083 Results-1547 Discussion-1823 Experimental Procedures-1898 Acknowledgements-118 References-2259 Figure Legends-897 1 Present address; Dept. of Bioscience, Tokyo University of Agriculture Sakuragaoka, Setagaya-ku, Tokyo 156-8502 Japan
46
Embed
Characterization and Functional Analysis of ABSCISIC ACID ... · Characterization and Functional Analysis of ABSCISIC ACID INSENSITIVE3-like Genes from Physcomitrella patens ... signaling
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
1
Characterization and Functional Analysis of ABSCISIC ACID INSENSITIVE3-like Genes from Physcomitrella patens Heather H. Marella*, Yoichi Sakata1*, and Ralph S. Quatrano2 Department of Biology, Washington University, 1 Brookings Dr., St. Louis, MO 63130, USA * These authors contributed equally to this work 2 Corresponding Author: Ralph Quatrano Address: Washington University, 1 Brookings Dr., Campus Box 1137, St. Louis, MO 63130 Email: [email protected] Telephone: 314-935-6850 Fax: 314-935-8137 Email of authors: Heather Marella- [email protected] Yoichi Sakata- [email protected] Running title: Physcomitrella patens ABI3
Word Count: Total-9875 Summary-250 Introduction-1083 Results-1547 Discussion-1823 Experimental Procedures-1898 Acknowledgements-118 References-2259 Figure Legends-897 1 Present address; Dept. of Bioscience, Tokyo University of Agriculture Sakuragaoka, Setagaya-ku, Tokyo 156-8502 Japan
2
SUMMARY
Although the moss Physcomitrella patens is known to respond to abscisic acid (ABA) by
activating gene expression, the transcriptional components involved have not been
characterized. Initially, we used the ABA-responsive Em promoter from wheat linked to
β-glucuronidase (GUS) to determine if ABI3/VP1, transcriptional regulators in the ABA
signaling pathway in angiosperms, were similarly active in the ABA response of P.
patens. We show by particle bombardment that ABI3 and VP1 affect Em-GUS
expression in P. patens in a manner similar to angiosperms. We also show the
involvement of ABI1 in the pathway, utilizing the abi1-1 mutant allele. We isolated
three ABI3-like genes from P. patens. Using an Em-like ABA-responsive promoter from
P. patens (PpLea1), we demonstrate that PpABI3A, only in the presence of ABA,
strongly enhances PpLea1-GUS expression in P. patens. PpABI3A also enhances ABA-
induced Em-GUS expression in P. patens. In barley aleurone, PpABI3A transactivates
Em-GUS but to a lesser extent than VP1 and ABI3. PpABI3A:GFP is localized to the
nucleus of both protonemal cells and barley aleurone indicating that the nuclear
localization signals are conserved. We show that at least a part of the inability of
PpABI3A to fully complement the phenotypes of the Arabidopsis abi3-6 mutant is due to
a weak interaction between PpABI3A and the bZIP transcription factor ABI5, as assayed
functionally in barley aleurone and physically in the yeast-two-hybrid assay. Our data
clearly demonstrate that P. patens will be useful for comparative structural and functional
studies of components in the ABA response pathway such as ABI3.
3
INTRODUCTION
The phytohormone abscisic acid (ABA) not only regulates processes occurring during
seed development (e.g. desiccation tolerance), but also controls processes associated with
responses to water stress during vegetative development of seed plants (e.g. stomatal
opening and closing). Since ABA is found in most land plants (Finkelstein and Rock,
2002), and has demonstrated physiological (Goode et al., 1993; Minami et al., 2003;
Minami et al., 2005; Werner et al., 1991) and molecular responses in non-seed plants
such as mosses (Knight et al., 1995), we have decided to take a comparative approach to
determine the evolution of this response pathway as well as the role of specific regulatory
proteins and protein domains that may be conserved.
Genetic approaches in Arabidopsis have been primarily responsible for
identifying several components involved in the ABA response pathway in seed plants.
Arabidopsis ABA-insensitive (abi) mutants were isolated by their ability to germinate in
the presence of ABA (Brocard-Gifford et al., 2004; Finkelstein, 1994; Koornneef et al.,
1984) and have been extensively characterized. The abi genes that have been cloned
revealed a diverse set of proteins including ABI1 and ABI2 which encode type 2C protein
phosphatases (PP2Cs) and are ABA signaling intermediates that act as negative
regulators (Gosti et al., 1999; Leung et al., 1994; Leung et al., 1997). The ABI3
(Giraudat et al., 1992), ABI4 (Finkelstein et al., 1998) and ABI5 (Finkelstein and Lynch,
2000) genes each encode a different type of transcriptional regulator, while ABI8
represents a novel plant specific protein (Brocard-Gifford et al., 2004). We will take a
comparative approach to characterize the ABA response in Physcomitrella patens and
will initially focus on the plant specific transcriptional regulator ABI3 from Arabidopsis,
4
and its ortholog from maize VP1 (Giraudat et al., 1992; McCarty et al., 1991) and
determine if a similar regulator is found in P. patens.
ABI3/VP1-like genes have been found in various seed plants (Bobb et al., 1995;
Chandler and Bartels, 1997; Footitt et al., 2003; Hattori et al., 1994; Lazarova et al.,
2002; Rohde et al., 2002; Shiota et al., 1998), and regulate a set of proteins expressed
during the later stages of seed development. One such gene, the Em gene (Marcotte et
al., 1989), requires both ABA and ABI3/VP1 for expression (Bies-Etheve et al., 1999;
McCarty et al., 1991; Vasil et al., 1995). All of the ABI3/VP1-like genes cloned from
seed plants revealed highly conserved protein domains, designated A1, B1, B2 and B3,
starting from the N-terminal (Suzuki et al., 1997). In fact, maize VP1 can complement
the major phenotypes of the Arabidopsis abi3-6 mutant (Suzuki et al., 2001). The B3
domain has been shown to bind DNA in vitro (Suzuki et al., 1997), whereas the B1
domain is involved in the physical interaction with the bZIP transcription factor, ABI5
(Nakamura et al., 2001). The B2 domain has been shown to be responsible for the ABA-
dependent activation of ABA-regulated genes, like Em, through the ABA-responsive
element (ABRE) (Bies-Etheve et al., 1999; Ezcurra et al., 2000; Hill et al., 1996), and
facilitates the interaction with bZIP transcription factors (Hill et al., 1996) such as ABI5.
However, the mechanism by which this domain functions in seed plants remains to be
elucidated. Until recently, ABI3/VP1 was thought to function exclusively during seed
development, specifically as a component of the ABA signaling pathway involved in the
maturation and germination of seeds (Giraudat et al., 1992; McCarty et al., 1991;
Nambara et al., 1995; Nambara et al., 2000; Parcy et al., 1997). However, recent reports
have pointed out that ABI3 might have broader functions outside of the seed, such as
5
plastid development, flowering time, and outgrowth of axillary meristems (reviewed in
Rohde et al., 2000). These analyses also revealed a novel crosstalk between ABA and
auxin in seed germination and lateral root formation in Arabidopsis (Brady et al., 2003;
Suzuki et al., 2001). These data clearly indicate that ABI3/VP1 is not only involved in
the ABA-regulation of seed development and germination, but also has broader functions
in vegetative growth.
The ABA response pathway in the moss P. patens was demonstrated previously
using the ABA-responsive promoter of the wheat Em gene. The wheat Em promoter can
be activated by exogenous ABA in a transient assay using protonemal tissue as well as in
stable expression lines of P. patens (Knight et al., 1995). Also, the in vitro footprint of
proteins of P. patens on the Em promoter was identical to that of seed plants, indicating
that the moss transcriptional machinery recognizes the same promoter region as the seed
plant factors (Knight et al., 1995). This suggests that higher plants and P. patens share
common ABA regulatory components and as such will be suitable for a comparative
approach to elucidate the mechanism(s) involved and the evolution of the transcriptional
response to ABA. Furthermore, a homolog of the wheat Em gene from P. patens,
PpLEA1, has recently been described (Kamisugi and Cuming, 2005). Similar to the
wheat Em gene, the expression of PpLEA1 is highly inducible by ABA and is mediated
through an ACGT motif in the promoter, a common feature of ABA-inducible genes in
seed plants (Kamisugi and Cuming, 2005). This provides another comparative tool for
the study of the ABA response pathway in P. patens.
We have started to dissect the ABA-regulated transcriptional mechanism in P.
patens by characterizing the structure and function of ABI3-like genes. We show by
6
particle bombardment that the seed plant proteins, ABI1, ABI3, and VP1 can affect Em-
GUS expression in P. patens. We successfully cloned three P. patens cDNAs encoding
ABI3/VP1-like genes and tested their activity in both P. patens and barley aleurone cells
using ABA-responsive Group 1LEA promoters from P. patens and wheat (Em).
PpABI3A enhances ABA-induced PpLea1-GUS and Em-GUS expression in protonemal
tissue and in barley aleurone cells, similar to the response elicited by ABI3/VP1.
However, PpABI3A cannot significantly enhance GUS expression in the absence of ABA
unlike ABI3/VP1. We also demonstrate that PpABI3A is able to function in certain
cellular and molecular functions both in protonemal tissue and in aleurone cells. We
show that at least a part of the inability of PpABI3A to fully complement the molecular
response in the Arabidopsis abi3-6 mutant is due to a weak interaction between
PpABI3A and the bZIP transcription factor ABI5, as assayed functionally in barley
aleurone and physically in the yeast-two-hybrid assay. Our data clearly demonstrate that
P. patens will be useful for comparative structural and functional studies of components
in the ABA response pathway such as ABI3.
7
RESULTS
ABA transcriptional regulation and signaling in P. patens is similar to that of higher
plants
In order to determine if the seed plant proteins ABI1, VP1 and ABI3 can act in the ABA
induced gene expression pathway, we tested their function in P. patens protonemal tissue.
Co-bombardment of the reporter Em-GUS with either Ubi-VP1 or Ubi-ABI3
demonstrated that both VP1 and ABI3 were able to transactivate Em-GUS expression,
four- and ten-fold respectively (Figure 1A). This response to VP1 and ABI3, in the
absence of ABA, is similar to the effect each has in the seed plant ABA response
pathway. In Arabidopsis and barley, ABI3 and VP1 characteristically act downstream
from ABI1 in the ABA signaling pathway (Brady et al., 2003; Casaretto and Ho, 2003).
Predictably, over-expression of the dominant negative allele of ABI1, 35S-abi1-1
(Armstrong et al., 1995), with Em-GUS completely repressed induction of Em-GUS
expression by ABA (Figure 1B). Furthermore, Ubi-VP1 could overcome the repression
effect of abi1-1 placing VP1 also downstream from ABI1 in the ABA signaling pathway
(Figure 1B). These results suggest the involvement of molecules in P. patens with the
same or similar functions as the regulatory ABI3/VP1 proteins from angiosperms.
8
Figure 1
Seed plant ABA response factors function in P. patens.
A) Maize VP1 and Arabidopsis ABI3 transactivate Em-GUS expression in P. patens.
Em-GUS and Ubi-LUC were co-bombarded into P. patens protonemal tissue with or
without the effector constructs, Ubi-VP1 or Ubi-ABI3, using 0.2µg of each construct.
Bars indicate the relative GUS activities ±SE after 48h incubation (n=4).
B) Em-GUS expression in P. patens is repressed by abi1-1 and restored by VP1.
Em-GUS and Ubi-LUC were co-bombarded into P. patens protonemal tissue with or
without the effector constructs, 35S-abi1-1 and Ubi-VP1, using 0.2µg of each construct.
Bars indicate the relative GUS activities ±SE after 48h incubation with (black bars) or
without (white bars) 10µM ABA (n=4).
0
30
60
90
120
150
180
GU
S/L
UC
+ VP1 Em +abi1-1 +abi1-1
GUSEm 35S 3’
Reporter
Effector
abi1-135S NOS 3’
VP1Ubi NOS 3’
Em +VP1 +ABI30
10
20
30
40
50
60
GU
S/L
UC
GUSEm 35S 3’
Reporter
Effector
VP1Ubi NOS 3’
ABI3Ubi NOS 3’
A B
9
Identification of ABI3/VP1-like proteins from P. patens
By searching public EST and genomic databases, we cloned three PpABI3 genes. The
PpABI3A gene encodes a 658 amino acid product, while the PpABI3B gene and PpABI3C
gene encode 515 and 539 amino acid products, respectively. The domain structure of
PpABI3A, PpABI3B, PpABI3C, AtABI3, and VP1 is relatively conserved (Figure 2A).
However, PpABI3B is missing the acidic activation and the serine-rich domains, while
PpABI3C contains a serine-rich region, but lacks the activation domain (Figure 2A). The
position of three of the introns of the B3 domain is conserved across all species, but
PpABI3C has all five of the intron positions found in the higher plant ABI3/VP1 (Figure
2A).
The amino acid sequence alignment of PpABI3A, PpABI3B, PpABI3C, AtABI3,
and VP1 reveals that the B3 domain, underlined in blue, is the most highly conserved
with 50% identity in all five sequences (Figure 2B). The B1 domain, underlined in red,
and the B2 domain, underlined in green, have 23% and 40% identity in the five proteins,
respectively (Figure 2B). The P. patens PpABI3 sequences are more closely related to
each other than the higher plant sequences and share 34% identity over their entirety.
PpABI3A, PpABI3B and PpABI3C share higher identity in the conserved basic domains;
B1 is 68% identical, while B2 and B3 are 60% and 64% identical, respectively. These
data strongly suggest that higher plant VP1/ABI3 and P. patens ABI3A, ABI3B, and
ABI3C originated from a common ancestor gene.
10
Figure 2
Comparison of the PpABI3 proteins with Arabidopsis ABI3 and maize VP1.
A) Schematic representation of PpABI3A, PpABI3B, PpABI3C, ABI3 and VP1 proteins.
Conserved basic regions (B1-B3) and the Serine-rich regions (S-rich) are indicated by
boxes. Asterisks indicate the position of introns.
We thank Dr. Peter McCourt for the abi3-6 seeds, Dr. Andrew Cuming for the PpLea1-
GUS construct, and Dr. Tuan-hua David Ho and Dr. Jose Casaretto for providing DNA
constructs of Ubi-HvABI5, Ubi-GFP, and Ubi-LUC. We thank Jana Hakenjos, Julie
Thole and Paul Klueh for their technical assistance and Drs. Ho and Casaretto for their
help with the particle bombardment of barley aleurone cells. We thank Yoshinori Murai
and Takahiro Kawato for their technical assistance with the yeast-two-hybrid
experiments. Some of the sequence data were produced by the U.S. Department of
Energy Joint Genome Institute http://www.jgi.doe.gov/. This work was supported
by a JSPS Postdoctoral Fellowship for Research Abroad to Y.S. and by The Danforth
Foundation/Washington University to R.S.Q.
39
REFERENCES Altschul, S. F., Gish, W., Miller, W., Myers, E. W., and Lipman, D. J. (1990). Basic local alignment search tool. J. Mol. Biol. 215, 403-410. Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J., Zhang, Z., Miller, W., and Lipman, D. J. (1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25, 3389-3402. Armstrong, F., Leung, J., Grabov, A., Brearley, J., Giraudat, J., and Blatt, M. R. (1995). Sensitivity to abscisic acid of guard-cell K+ channels is suppressed by abi1-1, a mutant Arabidopsis gene encoding a putative protein phosphatase. Proc. Natl. Acad. Sci. USA 92, 9520-9524. Benfey, P. N., Ren, L., Chua, N. H. (1990). Tissue-specific expression from CaMV 35S enhancer subdomains in early stages of plant development. EMBO J. 9, 1677-1684. Bezanilla, M., Pan, A., and Quatrano, R. S. (2003). RNA interference in the moss Physcomitrella patens. Plant Physiol. 133, 470-474. Bies-Etheve, N., da Silva Conceicao, A., Giraudat, J., Koornneef, M., Leon-Kloosterziel, K., Valon, C., and Delseny, M. (1999). Importance of the B2 domain of the Arabidopsis ABI3 protein for Em and 2S albumin gene regulation. Plant Mol. Biol. 40, 1045-1054. Bobb, A. J., Eiben, H. G., and Bustos, M. M. (1995). PvAlf, an embryo-specific acidic transcriptional activator enhances gene expression from phaseolin and phytohemagglutinin promoters. Plant J. 8, 331-343. Brady, S. M., Sarkar, S. F., Bonetta, D., and McCourt, P. (2003). The ABSCISIC ACID INSENSITIVE 3 (ABI3) gene is modulated by farnesylation and is involved in auxin signaling and lateral root development in Arabidopsis. Plant J. 34, 67-75. Brocard-Gifford, I., Lynch, T. J., Garcia, M. E., Malhotra, B., and Finkelstein, R. R. (2004). The Arabidopsis thaliana ABSCISIC ACID-INSENSITIVE8 locus encodes a novel protein mediating abscisic acid and sugar responses essential for growth. Plant Cell 16, 406-421. Bruce, W. B., Christensen, A. H., Klein, T., Fromm, M., and Quail, P. H. (1989). Photoregulation of a phytochrome gene promoter from oat transferred into rice by particle bombardment. Proc. Natl. Acad. Sci. USA 86, 9692-9696. Carrera, E., and Prat, S. (1998). Expression of the Arabidopsis abi1-1 mutant allele inhibits proteinase inhibitor wound-induction in tomato. Plant J. 15, 765-771.
40
Casaretto, J., and Ho, T. H. (2003). The transcription factors HvABI5 and HvVP1 are required for the abscisic acid induction of gene expression in barley aleurone cells. Plant Cell 15, 271-284. Chandler, J. W., and Bartels, D. (1997). Structure and function of the vp1 gene homologue from the resurrection plant Craterostigma plantagineum Hochst. Mol. Gen. Genet. 256, 539-546. Christensen, A. H., and Quail, P. H. (1996). Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants. Transgenic Res. 5, 213-218. Clough, S. J., Bent, A. F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 16, 735-743. Ezcurra, I., Wycliffe, P., Nehlin, L., Ellerstrom, M., and Rask, L. (2000). Transactivation of the Brassica napus napin promoter by ABI3 requires interaction of the conserved B2 and B3 domains of ABI3 with different cis-elements: B2 mediates activation through an ABRE, whereas B3 interacts with an RY/G-box. Plant J. 24, 57-66. Finkelstein, R. R. (1994). Mutations at two new Arabidopsis ABA response loci are similar to the abi3 mutations. Plant J. 5, 765-771. Finkelstein, R. R., Wang, M. L., Lynch, T. J., Rao, S., and Goodman, H. M. (1998). The Arabidopsis abscisic acid response locus ABI4 encodes an APETALA 2 domain protein. Plant Cell 10, 1043-1054. Finkelstein, R. R., and Lynch, T. J. (2000). The Arabidopsis abscisic acid response gene ABI5 encodes a basic leucine zipper transcription factor. Plant Cell 12, 599-609. Finkelstein, R. R., and Rock, C. D. (2002). Abscisic acid biosynthesis and response. The Arabidopsis Book, eds. C. R. Somerville and E. M. Meyerowitz, American Society of Plant Biologists, pp.1-48. Footitt. S., Ingouff, M., Clapham, D., von Arnold, S. (2003). Expression of the viviparous 1 (Pavp1) and p34cdc2 protein kinase (cdc2Pa) genes during somatic embryogenesis in Norway spruce (Picea abies [L.] Karst). J. Exp. Bot. 54, 1711-1719. Frank, W., Ratnadewi, D., Reski, R. (2005). Physcomitrella patens is highly tolerant against drought, salt and osmotic stress. Planta 220, 384-394. Gampala, S. S., Finkelstein, R. R., Sun, S. S., and Rock, C. D. (2002). ABI5 interacts with abscisic acid signaling effectors in rice protoplasts. J. Biol. Chem. 277, 1689-1694.
41
Giraudat, J., Hauge, B. M., Valon, C., Smalle, J., Parcy, F., and Goodman, H. M. (1992). Isolation of the Arabidopsis ABI3 gene by positional cloning. Plant Cell 4, 1251-1261. Goode, J. A., Stead, A. D., and Duckett, J. G. (1993). Redifferentiation of moss protonemata: an experimental and immunofluorescence study of brood cell formation. Can. J. Bot. 71, 1510-1519. Gosti, F., Beaudoin, N., Serizet, C., Webb, A. A., Vartanian, N., Giraudat, J. (1999). ABI1 protein phosphatase 2C is a negative regulator of abscisic acid signaling. Plant Cell 11, 1897-1910. Grabov, A., Leung, J., Giraudat, J., and Blatt, M. R. (1997). Alteration of anion channel kinetics in wild-type and abi1-1 transgenic Nicotiana benthamiana guard cells by abscisic acid. Plant J. 12, 203-213. Hagenbeek, D., Quatrano, R. S., and Rock, C. D. (2000). Trivalent ions activate abscisic acid-inducible promoters through an ABI1-dependent pathway in rice protoplasts. Plant Physiol. 123, 1553-1560. Hattori, T., Terada, T., and Hamasuna, S. T. (1994). Sequence and functional analyses of the rice gene homologous to the maize Vp1. Plant Mol. Biol. 24, 805-810. Hill, A., Nantel, A., Rock, C. D., and Quatrano, R. S. (1996). A conserved domain of the viviparous-1 gene product enhances the DNA binding activity of the bZIP protein EmBP-1 and other transcription factors. J. Biol. Chem. 271, 3366-3374. Hobo, T., Kowyama, Y., and Hattori, T. (1999). A bZIP factor, TRAB1, interacts with VP1 and mediates abscisic acid-induced transcription. Proc. Natl. Acad. Sci. USA 96, 15348-15353. James, P., Halladay, J., and Craig, E. A. (1996). Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. Genetics 144, 1425-1436. Jarvis, P., Chen, L. J., Li, H., Peto, C. A., Fankhauser, C., and Chory, J. (1998). An Arabidopsis mutant defective in the plastid general protein import apparatus. Science 282, 100-103. Jones, H. D., Kurup, S., Peters, N. C., and Holdsworth, M. J. (2000). Identification and analysis of proteins that interact with the Avena fatua homologue of the maize transcription factor VIVIPAROUS 1. Plant J. 21, 133-142. Kamisugi, Y. and Cuming, A.C. (2005). The evolution of the Abscisic acid-response in land plants: comparative analysis of Group 1 LEA gene expression in moss and cereals. Plant Mol. Biol. 59, 723-737.
42
Knight, C. D., Sehgal, A., Atwal, K., Wallace, J. C., Cove, D. J., Coates, D., Quatrano, R. S., Bahadur, S., Stockley, P. G., and Cuming, A. C. (1995). Molecular responses to abscisic acid and stress are conserved between moss and cereals. Plant Cell 7, 499-506. Koornneef, M., Reuling, G., and Karssen, C. M. (1984). The isolation and characterization of abscisic acid-insensitive mutants of Arabidopsis thaliana. Physiol. Plant 61, 377-383. Kurup, S., Jones, H. D., and Holdsworth, M. J. (2000). Interactions of the developmental regulator ABI3 with proteins identified from developing Arabidopsis seeds. Plant J. 21, 143-155. Lanahan, M. B., Ho, T. H., Rogers, S. W., and Rogers, J. C. (1992). A gibberellin response complex in cereal alpha-amylase gene promoters. Plant Cell 4, 203-211. Lara, P., Onate-Sanchez, L., Abraham, Z., Ferrandiz, C., Diaz, I., Carbonero, P., and Vicente-Carbajosa, J. (2003). Synergistic activation of seed storage protein gene expression in Arabidopsis by ABI3 and two bZIPs related to OPAQUE2. J. Biol. Chem. 278, 21003-21011. Lazarova, G., Zeng, Y., and Kermode, A. R. (2002). Cloning and expression of an ABSCISIC ACID-INSENSITIVE 3 (ABI3) gene homologue of yellow-cedar (Chamaecyparis nootkatensis). J. Exp. Bot. 53, 1219-1221. Leung, J., Bouvier-Durand, M., Morris, P.C., Guerrier, D., Chefdor, F., and Giraudat, J. (1994). Arabidopsis ABA response gene ABI1: features of a calcium-modulated protein phosphatase. Science 264, 1448-1452. Leung, J., Merlot, S., and Giraudat, J. (1997). The Arabidopsis ABSCISIC ACID- INSENSITIVE2 (ABI2) and ABI1 genes encode homologous protein phosphatases 2C involved in abscisic acid signal transduction. Plant Cell 9, 759-771. Marcotte, W. R., Jr., Russell, S. H., and Quatrano, R. S. (1989). Abscisic acid-responsive sequences from the em gene of wheat. Plant Cell 1, 969-976. McCarty, D. R., Hattori, T., Carson, C. B., Vasil, V., Lazar, M., and Vasil, I. K. (1991). The Viviparous-1 developmental gene of maize encodes a novel transcriptional activator. Cell 66, 895-905. Minami, A., Nagao, M., Arakawa, K., Fujikawa, S., and Takezawa, D. (2003). Abscisic acid-induced freezing tolerance in the moss Physcomitrella patens is accompanied by increased expression of stress-related genes. J. Plant Physiol. 160, 475-483. Minami, A., Nagao, M., Ikegami, K., Koshiba, T., Arakawa, K., Fujikawa, S.,
43
Takezawa, D. (2005). Cold acclimation in bryophytes: low-temperature-induced freezing tolerance in Physcomitrella patens is associated with increases in expression levels of stress-related genes but not with increase in level of endogenous abscisic acid. Planta 220, 414-423. Nakamura, S., Lynch, T. J., and Finkelstein, R .R. (2001). Physical interactions between ABA response loci of Arabidopsis. Plant J. 26, 627-635. Nambara, E., Keith, K., McCourt, P., and Naito, S. (1994). Isolation of an internal deletion mutant of the Arabidopsis thaliana ABI3 gene. Plant Cell Physiol. 35, 509-513. Nambara, E., Keith, K., McCourt, P., and Naito, S. (1995). A regulatory role for the ABI3 gene in the establishment of embryo maturation in Arabidopsis thaliana. Development 121, 629-636. Nambara, E., Hayama, R., Tsuchiya, Y., Nishimura, M., Kawaide, H., Kamiya, Y., and Naito, S. (2000). The role of ABI3 and FUS3 loci in Arabidopsis thaliana on phase transition from late embryo development to germination. Dev. Biol. 220, 412-423. Nambara, E., Suzuki, M., Abrams, S., McCarty, D. R., Kamiya, Y., and McCourt, P. (2002).A screen for genes that function in abscisic acid signaling in Arabidopsis thaliana. Genetics 161, 1247-1255. Nishiyama, T., Fujita, T., Shin, I.T., Seki, M., Nishide, H., Uchiyama, I., Kamiya, A., Carninci, P., Hayashizaki, Y., Shinozaki, K., Kohara, Y., and Hasebe, M. (2003). Comparative genomics of Physcomitrella patens gametophytic transcriptome and Arabidopsis thaliana: implication for land plant evolution. Proc. Natl. Acad. Sci. USA 100, 8007-8012. Notredame, C., Higgins, D., and Heringa, J. (2000). T-Coffee: a novel method for multiple sequence alignments. J. Mol. Biol. 302, 205-217. Parcy, F., Valon, C., Raynal, M., Gaubier-Comella, P., Delseny, M., Giraudat, J. (1994). Regulation of gene expression programs during Arabidopsis seed development: roles of the ABI3 locus and of endogenous abscisic acid. Plant Cell 6, 1567-1582. Parcy, F., Valon, C., Kohara, A., Misera, S., and Giraudat, J. (1997). The ABSCISIC ACID-INSENSITIVE3, FUSCA3, and LEAFY COTYLEDON1 loci act in concert to control multiple aspects of Arabidopsis seed development. Plant Cell 9, 1265-1277. Rohde, A., Kurup, S., and Holdswoth, M. (2000). ABI3 emerges from the seed. Trends Plant Sci. 5, 418-419. Rohde, A., Prinsen, E., De Rycke, R., Engler, G., Van Montagu, M., and Boerjan, W. (2002). PtABI3 impinges on the growth and differentiation of embryonic leaves during bud set in poplar. Plant Cell 14, 1885-1901.
44
Sambrook, J., and Russell, D.W. (2001). Molecular Cloning: A Laboratory Manual. (Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press). Schultz, T. F., Medina, J., Hill, A., and Quatrano, R. S. (1998). 14-3-3 proteins are part of an abscisic acid-VIVIPAROUS1 (VP1) response complex in the Em promoter and interact with VP1 and EmBP1. Plant Cell 10, 837-847. Shen, Q.J., Casaretto, J.A., Zhang, P., and Ho, T-H.D. (2004). Functional definition of ABA-response complexes: the promoter units necessary and sufficient for ABA induction of gene expression in barley (Hordeum vulgare L.). Plant Mol. Biol. 54, 111-124. Shen, Q., Uknes, S.J., and Ho, T. H. (1993). Hormone response complex in a novel abscisic acid and cycloheximide-inducible barley gene. J. Biol. Chem. 268, 23652-23660. Shiota, H., Satoh, R., Watabe, K., Harada, H., and Kamada, H. (1998). C-ABI3, the carrot homologue of the Arabidopsis ABI3, is expressed during both zygotic and somatic embryogenesis and functions in the regulation of embryo-specific ABA-inducible genes. Plant Cell Physiol. 39, 1184-1193. Sunilkumar, G., Mohr, L., Lopata-Finch, E., Emani, C., Rathore, K. S. (2002). Developmental and tissue-specific expression of CaMV 35S promoter in cotton as revealed by GFP. Plant Mol Biol. 50, 463-474. Suzuki, M., Kao, C. Y., and McCarty, D. R. (1997). The conserved B3 domain of VIVIPAROUS1 has a cooperative DNA binding activity. Plant Cell 9, 799-807. Suzuki, M., Kao, C.Y., Cocciolone, S., and McCarty, D. R. (2001). Maize VP1 complements Arabidopsis abi3 and confers a novel ABA/auxin interaction in roots. Plant J. 28, 409-418. Terada, R., Shimamoto, K. (1990). Expression of CaMV 35S-Gus gene in transgenic rice plants. Mol. Gen. Genet. 220, 389-392. Vasil, V., Marcotte, W. R., Jr., Rosenkrans, L., Cocciolone, S. M., Vasil, I. K., Quatrano, R. S., and McCarty, D. R. (1995). Overlap of Viviparous1 (VP1) and abscisic acid response elements in the Em promoter: G-box elements are sufficient but not necessary for VP1 transactivation. Plant Cell 7, 1511-1518. Werner, O., Ros Espin, R. M., Bopp, M., and Atzorn, R. (1991). Abscisic-acid-induced drought tolerance in Funaria hygrometrica Hedw. Planta 186, 99–103. Zeng, Y., Kermode, A. R. (2004). A gymnosperm ABI3 gene functions in a severe abscisic acid-insensitive mutant of Arabidopsis (abi3-6) to restore the wild-type
45
phenotype and demonstrates a strong synergistic effect with sugar in the inhibition of post-germinative growth. Plant Mol. Biol. 56, 731-746. Zentella, R., Yamauchi, D., and Ho, T. H. (2002). Molecular dissection of the gibberellin/abscisic acid signaling pathways by transiently expressed RNA interference in barley aleurone cells. Plant Cell 14, 2289-2301. Zhang, X., Garreton, V., Chua, N. H. (2005). The AIP2 E3 ligase acts as a novel negative regulator of ABA signaling by promoting ABI3 degradation. Genes Dev. 1, 1532-1543.
46
Table 1. AtABI3 regulated genes grouped by co-regulator for RT-PCR analysis.