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DNA Sqirence-The loirrnnl of Seqirencing and Mopping, Vol. 8 (1-Z), pp. 109-112 Reprints available directly from the publisher Photocopying permitted by license only
0 1997 OI'A (Overseas Publishers Association) Amsterdam B.V. Published in The Netherlands
by Hanvood Academic Publishers Printed in India
Short Communication
Nucleotide Sequence of Grapevine (Vitis vinifera) cDNA Similar to SNAP Proteins S. MATSUMOTOa,*, IAN B. DRYb and M. THOMASb
'Department of Biology, Faculty of Education, Gifu University, Gifu 501721, Japan; bCSIRO Division of Horticulture, GPO Box 350, Adelaide, SA 5001, Australia
Database Accession No.: AB001375
(Received 28 Februa y 1997)
A cDNA clone (VSl) homologous to SNAP proteins was isolated from a grapevine cDNA library. The cDNA insert was 1167 bp long and contained a single open reading frame coding for 289 amino acids. The amino acid sequence of VS1 shows similarity (35%-45%) to SNAP proteins from various sources.
Keywords: cDNA, grapevine, SNAP, Vitis vinifera
SNAPs (soluble NSF attachment proteins) are cytoplasmic proteins that bind to membranes via specific receptors referred to as v- and t- SNAP receptors (SNARES). SNAPs and NSF (N-ethylmaleimide-sensitive factor), a protein that is required for membrane fusion reactions, are essential for membrane traffic (Beckers et al., 1989; Wilson et al., 1989).
A cDNA clone, named VS1, was obtained from a Vitis vinifera cv. Shiraz pre-veraison berry cDNA library. The nucleotide and deduced amino acid sequence of the cDNA insert is shown in Figure 1. The cDNA insert contains 867 bp of
coding region encoding 289 amino acids. The termination codon, TGA, is present at position 870-872, followed by the 3' untranslated region.
By comparing nucleotide sequence of the cDNA insert to the DNA (DDBJ, EMBL and GenBank) database rereased at Nov. or Dec. 1996, we found it showing high similarity with cDNA clones TAP0070 from Arabidopsis thaliana and S3606-1A from Oryza sativa. Both cDNA clones are ESTs (expressed sequence tags), and contain partial 5' end cDNA sequences. As the first 88 and 103 deduced amino acids of the VS1 show 65% and 60% sequence identity with that of TAP0070 and S3606_1A, respectively (Fig. 2), these 3 clones might be closely related. Since the clone TAP0070 possesses similarity to Bos Taurus SNAP protein, we searched the NRP (Swiss-Prot, PIR, Genpept and PDB) data base using deduced amino acid sequence of the VS1. The VS1 shows 45% and 44% sequence identity with Lorigo pealei and Bos prirnigenius taurus SNAP proteins (Fig. 3).
* Corresponding author. Tel.: +81-58-293-2257. Fax: +81-58-293-2207. E-mail [email protected].
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cc ATGGCGGATAATATACAGAGAGGAGAAGAGTTCGAGAATGCCGTTAGAAAGCGGAGAATGCCGTTAGAAGATCAGT M A D N I Q R G E E F E K K A E K K I S
2 62 20
GGCTGGGGATTGTTCGGTTCCAAATACGAATGCCGTTAGATGCTGCTGATTTCTACGATAAGGCTGCC G W G L F G S K Y E D A A D F Y D K A A
122 40
AACTGCTTCAAACTCGCCAAATCCTGGGACAGAGCTGGATCGACTTATGTCAAGTTGTCA N C F K L A K S W D R A G S T Y V K L S
182 60
ACTGTTATACAAAGCTCGGATAGCAAACATGAATGCCGTTAGCTGCACAATGCCGTTAGCTTATGCTGATGCTGGT T V I Q S S D S K H E A A Q A Y A D A G
242 80
CATT~CTATAAGAAAACATCTGCTAAAGAAGCCATTTCTTGCTTAGAACAAGCAGCATAT H C Y K K T S A K E A I S C L E Q A A Y
302 1 0 0
CTGTTTTTGGATAATGGGAGGTTCAACATGGCTGGAAAATATTACAAGGAAATTGCTGAA L F L D N G R F N M A G K Y Y K E I A E
362 1 2 0
TTATATGAGCTTGAACAAAACTTTGAGCAGGCTATAATCTACTTTG~GCAGCTGAT L Y E L E Q N F E Q A I I Y F E K A A D
422 1 4 0
ATTTATCAAAGTGAAGAAGCAACAACTGCTGCAAACCAGTGTAATGCAAAAGTTGCACAA I Y Q S E E A T T A A N Q C N A K V A Q
482 1 6 0
TTTGCTGCTCAACTAGAACAATATCAGAATGCCGTTAGGCAATGCCGTTATTCAAGACATAGGACGA F A A Q L E ’ Q Y Q K A I Q I Y E D I G R
542 1 8 0
CCATCACTAAACAATAATTTGCTGAAGTATGGAGTCAAGGGACATCTTCTTAACGCTGGC P S L N N N L L K Y G V K G H L L N A G
602 200
ATTTGTCAACTTTGCAAAGGCGATGTCGTTGCAATTACCAATGCCGTTATGCATTAGACCGATATCAG I C Q L C K G D V V A I T N A L D R Y Q
662 220
GAAATGGATCCAACTTTTTCAGGAACACGTGAATATAAATTACTTGTGGATTTGGCTGCT E M D P T F S G T R E Y K L L V D L A A
722 240
GCAGTTGATGAGGAAGATGTTGTGAAGTTCACTGATGCTGTTAAGGAATTTGATAGCATG A V D E E D V V K F T D A V K E F D S M
782 260
ACCCAACTGGATGCTTGGAAGACAACCCTTCTACTGAGGGTG~GAAGCTATCAAAGCC T Q L D A W K T T L L L R V K E A I K A
842 280
IlllAGAACTAGAGGAGGATGATCTCACCTGAGTCCTATTTCTCGGTTCCGTTTCAGGGTTT K E L E E D D L T *
902 289
TGTTCAGGATGATTTCTTTGTATAATGCCGTTATGCCGTTATGTCTGCCACACATTCACGATCTCCAA TTTTTTCTTTTTGCTTTCTTCAAATCTTCTTGATTTGGGTTTCAATCAGAATGTCTATGC TTGCATTTCTGAGGGTGAGGTTCTAATGCCGTTATGTGATCATTGTTTTCCTTGCTTGTATACACACA CCTACCATTTCTGAATTCTCAATGTCGGACTCAAGCG~TGATTGAATTT~TGGTT GTGCTTTTATGAAT-
962 1022 1082 1142 1167
FIGURE 1 Nucleotide and deduced amino acid sequences of VSI. The asterisk indicates the stop codon.
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Arabidopsis
Grape
Rice
Arabidopsis
Grape
Rice
10 20 30 40 50 MGDHLVRAEEFEKKAEKKLNGWGIFGSKYEDAADLLEKAANSYKLAKSWD I I I I I I I I I I I I I I l l I I I I I I I I I I I I I I I I I I I I I MADNIQRGEEFEKKAEKKISGWGLFGSKYEDAADFYDKAANCFKLAKSWD I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I MGDHEARGDDFEKKAEKKLSGWGIFGSKYEDAADLFDKGANSFKLAKNWS
60 70 80 90 100 QAGKAYLKLADCHLKSDSKHDAANAYAEAAKCYKKVGH
RAGSTWKLSTVIQSSDSKHEAAQAYADAGHCYKKTSAKEAISCLEQAAYLFL I I I I I I I I I I I I I I I l l1 I I I I I I l l RAAQVYIKIANCHLKGDSKHEAASAWEAANCYKKFSPQEAXQALNQXVDLFL
I I I l l I I I I I I I I l l I I I I I
FIGURE 2 Comparison of the amino acid sequences. The sequences presented are: Arabidopsis, cDNA clone TAP0070 (Accession No. F14363) from Arabidopsis thaliana (Cooke et ul., unpublished results); Grape, N-terminal region of VS1 from Vitis vinifcra; Rice, cDNA clone S3606JA (Accession No. D41242) from Oiyza sativa (Sasaki et al., unpublished results).
10 20 30 40 50 S q u i d MADN--EQKAMQLMQEAEKKLGSSKGFLSSLFGGSSKTEEAAELFVRAAN
G r a p e MADN--IQRGEEFEKKAEKKISGW-----GLFG--SKYEDAADFYDKAAN
B o v i n MDNSGKEAEAMALLAEAERKVKNSQSFFSGLFGGSSKIEEACEIYARAAN
I I I I I I l l 1 I l l I I I I I I l l .
I I I I I I I I I I I I I l l
60 7 0 80 90 100 S q u i d TFKMAKNWASAGQCFCKAAQLQLTLQSRHEAATHYVDAGNAFKKADPNES
G r a p e CFKLAKSWDRAGSTYVKLSTVIQSSDSKHEAAQAYADAGHCYKKTSAKEA
B o v i n MFKMAKNWSAAGSAFCQAAHVHLQLQSKHDAATCFVDAGNAFKKADPQEA
I I I I I I I I I I I I I I l l 1 I 1 I
I I I I I I l l I I I I I I I l l I I I I
110 120 130 140 150 S q u i d INCLLKAVEIYTDMGRFTIAAKHHITVAEIYENELADIEQAMSHYERAAD
G r a p e ISCLEQAAYLFLDNGRFNMAGKYYKEIAELYELE-QNFEQAIIYFEKAAD
B o v i n lNCLMRAlE lYTDMGRFTlAAKHHlS lAElYETELVDlEKAIAHYEQSAD
I l l I I I I I I I I I I I I I l l I l l 1
I l l I I l l 1 I I I I I I I I I l l I I I
160 1 7 0 180 190 200 S q u i d YYKGEESNSAANKCFLKVAQYSAQLEQYQKAIDIYEQVASNCMENALLKY
G r a p e IYQSEEATTAANQCNAKVAQFAAQLEQYQKAIQIYEDIGRPSLNNNLLKY
B o v i n YYKGEESNSSANKCLLKVAGYAAQLEQYQKAIDIYEQVGTNAMDSPLLKY
I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I
I I I I l l I l l I I I I I I I I I I I I I I I I l l 1
210 220 230 240 2 5 0 S q u i d SAKDHFFRAALCHMCN-DIVNAKLVIPKYEEMFPAFSDSRECNLLKSVLD
G r a p e GVKGHLLNAGICQLCKGDVVAITNALDRYQEMDPTFSGTREYKLLVDLAA
B o v i n SAKDYFFKAALCHFCI-DMLNAKLAVQKYEELFPAFSDSRECKRIKKLLE
I I I I I I I I I I I I I I I I
I I I I I I I I I l l I I I
260 2 7 0 280 290 S q u i d SMD--NVDEFTKAVADYDSISRLDQWLTMMFLRIKNQITEEVTDEMNLC
G r a p e AVDEEDVVKFTDAVKEFDSMTQLDAWKTTLLLRVKEAIKAKELEEDDLT
B o v i n AHEEQNVDSYTEAVKEYDSlSRLDQWLTTMLLRlKKTlQG---DEEDLR
I I I I I I I I I l l 1 I l l I I I
I I I I I I I I I I I I I I I I I I I I I I I
FIGURE 3 Comparison of the amino acid sequences. The sequences presented are: Squid, SNAP from Lorigo pealei (DeBello et a/., 1995); Grape, VS1 from Vitis viniferu; Bovin, &SNAP from Bos priniigenius Taurus (Whiteheart et a/., 1993). Gaps introduced to opti- mize alignments are presented with dashes.
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Moreover, the VS1 shows similarity with Bos primigenius taurus P-SNAP (Whiteheart et al., 1993) (44%), Homo sapiens a-SNAP (Accession No. U39412, Chen et al . , unpublished results) (43%), Saccharomyces cerevisiae vesicular-fusion protein secl7, which is functionally equivalent to mammalian a-SNAP protein (Griff et al., 1992) (%YO), Coprinus cinereus secl7-like protein (lse17, Accession No. U79186, Maida et al., unpublished results) (41%), Drosophila melanogaster soluble NSF attachment protein (Ordway et al., 1994) (40%), Xattus norvegicus a-soluble NSF attach- ment protein (Accession No. X89968, Mitchell et al., unpublished results) (43%). The calculated molecular weight (32,483 dalton) and the number of amino acids (289) are also very close to SNAP proteins from various sources (31,946-33,533 dalton and 285-298 amino acid). Although it is difficult to elucidate the function of the VS1 on the basis of sequence data, it seems that VS1 is SNAP homologue in plant.
Acknow Zedgements
We thank Dr. C. Davies for donating the grape- vine cDNA library and Mr. K. Kitahara for tech-
nical assistance. This research was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports and Science of Japan (Nos. 07660032 and 07238210).
References
Beckers, C. J. M., Block, M. R., Glick, 8. S., Rothman, J. E. and Balch, W. E. (1989) ”Vesicular transport between the endo- plasmic reticulum and the Golgi stack requires the NEM- sensitive fusion protein,” Nature, 339,397-398.
Debello, W. M., OConner, V., Dresbach, T., Whiteheart, S. W., Wang, S. S.-H., Schweizer, F. E., Betz, H., Rothman, J. E. and Augustine, G. J. (1995) ”SNAP-mediated protein-protein interactions essential for neurotransmitter release,” Nature,
Griff, I. C., Schekman, R., Rothman, J. E. and Kaiser, C. A. (1992) “The yeast SEC27 gene product is functionally equivalent to mammalian a-SNAP protein,” J. Bid . Chem.,
Ordway, R. W., Pallanck, L. and Ganetzky, B. (1994) ”Neurally expressed Drosophila genes encoding homolo- gous of the NSF and SNAP secretory proteins,” Proc. Natl. Acad. Sci. USA, 91,5715-5719.
Whiteheart, S. W., Griff, I. C., Brunner, M., Clary, D. O., Mayer, T., Buhrow, S. A. and Rothman, J. E. (1993) “SNAP family of NSF attachment proteins includes a brain-specific isoform,” Nature, 362,353-355.
Wilson, D. W., Wilcox, C. A., Flynn, G. C., Chen, E., Kuang, W.-J., Henzel, W. J., Block, M. R., Ullrich, A. and Rothman, J. E. (1989) “A fusion protein required for vesicle-mediated transport in both mammalian cells and yeast,” Nature, 339, 355-359.
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