14-3-3 Proteins are components of the transcription complex of the ATEM1 promoter in Arabidopsis.
The AtEm1 and AtEm6 gene products accumulate exclusively in embryos during Arabidopsis seed maturation. The transcription factor ABI3 and the phytohormone abscisic acid are required for normal expression of both genes. However, the expression of these genes occurs in extremely small embryos limiting the availability of tissue to directly study DNA-protein interactions. We generated callus lines derived from embryos to determine if the regulation of Em expression was similar to wild type embryos. Expression of AtEm1 and AtEm6 was strongly induced by abscisic acid in callus derived from wild type embryos, but not in embryo callus derived from ABI3 mutant embryos (abi3-6). Epitopes to 14-3-3 proteins were found in complexes with the AtEm1 promoter in mobility shift experiments using nuclear extracts derived from both wild type and abi3-6 calli. Using phosphorylated peptides that bind to 14-3-3 proteins, we show that 14-3-3 proteins are required for the maintenance of the transcriptional complex generated in nuclear extracts. Chromatin immunoprecipitation experiments using a 14-3-3 antibody display the expected 241-bp band from the AtEm1 promoter. Hence, 14-3-3 proteins are physically present in the AtEm1 transcriptional complex in vivo and are required for the maintenance of the transcriptional complex in vitro.
del Viso
Florencia
F
Department of Biology, Washington University, St Louis, MO 63130, USA.
Casaretto
Jose A
JA
Quatrano
Ralph S
RS
D048948:14-3-3 Proteins / Q000235:genetics / Q000378:metabolism*; D000040:Abscisic Acid / Q000494:pharmacology; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D047369:Chromatin Immunoprecipitation; D024202:Electrophoretic Mobility Shift Assay; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000235:genetics; D017343:Genes, Plant / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics
D048948:14-3-3 Proteins; C091601:ABI3 protein, Arabidopsis; C509632:ATEM6 protein, Arabidopsis; D029681:Arabidopsis Proteins; D014157:Transcription Factors; D000040:Abscisic Acid
Plant J. 2002 May;30(3):373-83
12000684
BMC Bioinformatics. 2003 Oct 29;4:52
14583099
Plant J. 1996 Oct;10(4):655-61
8893542
Plant J. 2002 Nov;32(3):317-28
12410810
Plant Cell. 1992 Oct;4(10):1251-61
1359917
Plant Cell. 2000 Jan;12(1):35-52
10634906
Plant Cell. 1998 May;10(5):837-47
9596641
Plant J. 2007 Jan;49(2):289-301
17241451
Trends Plant Sci. 2003 May;8(5):218-23
12758039
Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11490-4
1454838
J Cell Biochem. 2002;87(2):194-207
12244572
Biochem J. 2005 May 15;388(Pt 1):151-7
15631617
Plant Cell. 1993 Oct;5(10):1401-10
8281041
Plant Mol Biol. 1999 Jan;39(1):117-28
10080714
Plant Cell. 1990 Jun;2(6):503-12
2152172
Plant Physiol. 1996 Jan;110(1):249-257
12226181
J Exp Bot. 2004 Jan;55(394):77-87
14676287
Plant Cell. 1997 Aug;9(8):1265-77
9286105
Plant Cell. 1994 Nov;6(11):1567-82
7827492
Development. 2003 Dec;130(24):6065-73
14597573
Plant Cell. 1994 Apr;6(4):501-10
8205002
Plant Cell Physiol. 2005 Mar;46(3):399-406
15695450
Plant J. 2002 Dec;32(5):831-43
12472697
Plant Physiol. 2000 Dec;124(4):1752-65
11115891
Planta. 1994;192(3):384-94
7764404
Mol Gen Genet. 1993 Apr;238(3):409-18
8492809
Plant Physiol. 1994 Dec;106(4):1593-604
7846163
Eur J Biochem. 1979 Sep;99(2):217-24
499199
Plant Cell. 2006 Jul;18(7):1642-51
16731585
Science. 1990 Oct 12;250(4978):267-71
2145628
Annu Rev Plant Physiol Plant Mol Biol. 1996 Jun;47:49-73
15012282
J Exp Bot. 2000 Jul;51(348):1211-20
10937696
Plant Physiol. 1993 Aug;102(4):1185-1191
12231895
Plant Physiol. 2006 Feb;140(2):647-60
16407442
Plant Cell Physiol. 1994 Apr;35(3):509-13
8055176
Planta. 2004 May;219(1):158-66
14767767
Plant J. 2005 Dec;44(6):1001-9
16359392
Plant Mol Biol. 1995 Dec;29(5):1015-26
8555445
Plant Cell. 1992 Oct;4(10):1295-307
1446170