Classic FYVE zinc-finger domains recognize the phosphoinositide signal PtdIns3P and share the basic (R/K)1(R/K)HHCR6 (single-letter amino acid codes) consensus sequence. This domain is present in predicted PtdIns3P 5-kinases and lipases from Arabidopsis thaliana. Other Arabidopsis proteins, named PRAF, consist of a pleckstrin homology (PH) domain, a regulator of chromosome condensation (RCC1) guanine nucleotide exchange factor repeat domain, and a variant FYVE domain containing an Asn residue and a Tyr residue at positions corresponding to the PtdIns3P-interacting His4 and Arg6 of the basic motif. Dot-blot and liposome-binding assays were used in vitro to examine the phospholipid-binding ability of isolated PRAF domains. Whereas the PH domain preferentially bound PtdIns(4,5)P2, the variant FYVE domain showed a weaker charge-dependent binding of phosphoinositides. In contrast, specificity for PtdIns3P was obtained by mutagenic conversion of the variant into a classic FYVE domain (Asn4,Tyr6 → His4,Arg6). Separate substitutions of the variant residues were not sufficient to impose preferential binding of PtdIns3P, suggesting a co-operative effect of these residues in binding. A biochemical function for PRAF was indicated by its ability to catalyse guanine nucleotide exchange on some of the small GTPases of the Rab family, permitting a discussion of the biological roles of plant FYVE proteins and their regulation by phosphoinositides.
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October 2001
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Research Article|
September 24 2001
FYVE zinc-finger proteins in the plant model Arabidopsis thaliana: identification of PtdIns3P-binding residues by comparison of classic and variant FYVE domains
Rikke B. JENSEN;
Rikke B. JENSEN
Institute of Molecular Biology, University of Copenhagen, Øster Farimagsgade 2A, 1353 Copenhagen K, Denmark
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Tanja LACOUR;
Tanja LACOUR
Institute of Molecular Biology, University of Copenhagen, Øster Farimagsgade 2A, 1353 Copenhagen K, Denmark
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Jakob ALBRETHSEN;
Jakob ALBRETHSEN
Institute of Molecular Biology, University of Copenhagen, Øster Farimagsgade 2A, 1353 Copenhagen K, Denmark
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Michael NIELSEN;
Michael NIELSEN
Institute of Molecular Biology, University of Copenhagen, Øster Farimagsgade 2A, 1353 Copenhagen K, Denmark
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Karen SKRIVER
Karen SKRIVER
1
Institute of Molecular Biology, University of Copenhagen, Øster Farimagsgade 2A, 1353 Copenhagen K, Denmark
1To whom correspondence should be addressed (e-mail ks@apk.molbio.ku.dk).
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Publisher: Portland Press Ltd
Received:
December 19 2000
Revision Received:
June 01 2001
Accepted:
July 31 2001
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2001
2001
Biochem J (2001) 359 (1): 165–173.
Article history
Received:
December 19 2000
Revision Received:
June 01 2001
Accepted:
July 31 2001
Citation
Rikke B. JENSEN, Tanja LACOUR, Jakob ALBRETHSEN, Michael NIELSEN, Karen SKRIVER; FYVE zinc-finger proteins in the plant model Arabidopsis thaliana: identification of PtdIns3P-binding residues by comparison of classic and variant FYVE domains. Biochem J 1 October 2001; 359 (1): 165–173. doi: https://doi.org/10.1042/bj3590165
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