Nucleic Acids Research, Vol. 18, No. 10 3049

©) 1990 Oxford University Press

Nucleotide sequence of the yeast UGA1 GABA transaminase

gene

encoding

Bruno Andre and Jean-Claude Jauniaux Laboratoire de Microbiologie, Faculte des Sciences, Universite Libre de Bruxelles, Campus de la Plaine CP244, Bd du Triomphe, B-1050 Bruxelles, Belgium Submitted April 18, 1990 The UGA I gene of Saccharomyces cerevisiae encodes 4-aminobutyrate:2-oxoglutarate aminotransferase (GABA transaminase) (EC 2.6.1.19), the first enzyme in the catabolism of 4-aminobutyrate (GABA) (1). A 7-kb BamHI-BamHI DNA fragment was isolated from a plasmid library representing the genome of the yeast E 1278b-strain for it's ability to restore growth to ugal mutant cells on a medium containing GABA as sole nitrogen source. The cloned DNA fragment was shown to direct integration to the UGAI chromosomal locus. The UGAI gene was subcloned on a 3.2-kb Smal-EcoRI DNA segment. The sequence of the UGAI gene and the deduced polypeptide are shown below. The UGAI predicted amino-acid sequence (471 aa; MW 52792) shares high similarities with yeast (2), rat (3) and human (3) ornithine aminotransferases, but only weak or no apparent similarities with the other aminotransferases thus far sequenced.

EMBL accession no. X52600 TAACTAAGAACA M

S

I

C

E

Q

Y

Y

P

E

E

P

T

K

P

GTTAAGACCGAG T V K T E

S

I

P

G

P

E

S

Q

K

Q

L

K

E

L

G

E

60 20

,GTTTTTGACACA V F D T

120 40

AGACCAGCATATTTTTTGGCTGATTATGAGAAATCTTTAGGCAACTAT. ATCACTGATGTG R P A Y F L A D Y E K S L G N Y I T D V

180 60

GATGGGAACACATATTTGGATTTGTATGCCCAAATCTCTTCAATTGCA ,CTTGGTTATAAC D G N T Y L D L Y A Q I S S I A L G Y N

240 80

AACCCTGCTTTGATCAAGGCAGCACAATCACCAGAAATGATCCGTGCT TTGGTCGACCGT N P A L I K A A Q S P E M I R A L V D R

300 100

CCTGCCTTAGGTAACTTCCCATCTAAGGATTTAGACAAGATATTGAAG ,CAAATATTGAAA P A L G N F P S K D L D K I L K Q I L K

360 120

TCTGCGCCAAAGGGTCAAGATCACGTCTGGTCAGGGCTTTCCGGTGCA ,GATGCCAATGAA S A P K G Q D H V W S G L S G A D A N E

420 140

TTAGCGTTCAAGGCTGCCTTTATTTATTACCGTGCCAAACAAAGGGGC' 'TATGATGCCGAT

480 160

L

A

F

K

A

A

F

I

Y

Y

R

A

K

Q

R

G

Y

D

A

D

TTTTCTGAAAAGGAAAACTTGTCTGTCATGGACAATGACGCCCCTGGC GCCCCTCATCTT F

S

E

K

E

N

L

S

V

M

D

N

D

A

P

G

A

P

H

L

GCCGTACTATCGTTCAAGAGAGCGTTCCACGGTAGATTGTTTGCCTCCGGTTCCACAACT

ACKNOWLEDGEMENTS

A

V

L

S

F

K

R

A

F

H

G

R

L

F

A

S

G

S

T

T

TGTTCTAAACCAATTCACAAGTTGGATTTCCCAGCCTTCCACTGGCCTCATGCTGAGTAT

We wish to thank S.Vissers for having provided the cloned UGAI gene and C.Pirlot for her contribution in chromosomal targeting and DNA sequencing experiments. This work was supported by contracts FRFC 4513.88 and ARC-SPPS 86/91-88. A.B. is recipient of a fellowship from the Van Buuren Foundation.

C

1. Ramos,F., El Guezzar,M., Grenson,M. and Wiame,J.-M. (1985) Eur. J.

Biochem. 149, 401-404. 2. Degols,G. (1987) Eur. J. Biochem. 169, 193-200. 3. Inana,G., Totsuka,S., Redmond,M., Dougherty,T., Nagle,J., Shiono,T., Ohura,T., Kominami,E. and Katunuma,N. (1985) Proc. Natl. Acad. Sci. USA 83, 1203-1207. 4. Kuramitsu,S., Ogawa,T., Ogawa,H. and Kagamiyama,H. (1985) J. Biol. Chem. (Tokyo) 97, 993-999.

S

K

P

I

H

K

L

D

F

P

A

F

H

W

P

H

A

E

Y

CCATCTTACCAATACCCATTAGATGAAAATTCTGATGCAAACCGTAAAGAGGATGACCGT P

S

Y

Q

Y

P

L

D

E

N

S

D

A

N

R

K

E

D

D

R

TGCTTGGCCATTGTTGAAGAATTAATCAAAACCTGGTCTATTCCAGTTGCTGCCTTGATC C

L

A

I

V

E

E

L

I

K

T

W

S

I

P

V

A

A

L

I

ATCGAACCAATTCAATCTGAGGGAGGTGATAACCACGCTTCTAAGTATTTCTTACAAAAG I

REFERENCES

-1

E

P

I

Q

S

E

G

G

D

N

H

A

S

K

Y

F

L

Q

K

CTAAGAGACATTACCTTGAAGTATAACGTTGTCTACATCATAGATGAAGTGCAAACAGGT L

R

D

I

T

L

K

Y

N

V

V

Y

I

I

D

E

V

Q

T

G

GTCGGAGCCACCGGTAAGCTATGGTGTCATGAGTACGCCGATATTCAACCACCTGTGGAT V

G

A

T

G

K

L

W

C

H

E

Y

A

D

I

Q

P

P

V

D

TTAGTGACCTTTTCCAAGAAATTCCAAAGTGCAGGATATTTCTTCCACGACCCTAAATTC L V T F S K K F Q S A G Y F F H D P K F I

P

N

K

P

Y

R

Q

F

N

T

W

C

G

E

P

A

R

M

I

ATTGCAGGTGCCATTGGACAGGAAATCTCCGACAAGAAGTTGACTGAACAATGTTCAAGA I A G A I G Q E I S D K K L T E Q C S R

540 180 600 200 660 220

720 240 780 260 840 280 900 300 960 320

1020 340 1080 360 1140 380

V

G

D

Y

L

F

K

K

L

E

G

L

Q

K

K

Y

P

E

N

F

1200 400

Q

N

L

R

G

K

G

R

G

T

F

I

A

W

D

L

P

T

G

E

1260 420

AAGAGAGACTTACTATTGAAGAAATTGAAGTTGAATGGTTGCAACGTTGGTGGATGTGCA K R D L L L K K L K L N G C N V G G C A

1320 440

F

1380 460

ATTGAAGCATTAGCCAAATCAGTTAATGAATTATGATTGTATATTAGCGAGGCCGTGTCA I E A L A K S V N E L

1440 471

CGTTCCCTTTTTTGGCCTTTATATATGTGTATCTTGTTATAATTTTAAAGAAAAGTTTTA TATTACTCATCAGTTGTGTGTGAAAAATATGGGCGGGTATTCACAATAGGAANNNAAAAA TGGGTTTGGTGACTGCAGTAAAGACGTGCTAAAAAAAAAAGTAGTTTTTATCTGAACCTT AAATATATCATTTATGAGTGTATATATTATACAAACGCAATAACAGTGGCTCCCGCCTCC ACCTCCGCCTGCTACTAGTAATCTACCAGTAATTGTGTTTGTTTTGGGAAAGATCGTTCG CTTACTCACCGAACAACTTTCTGACTCTCTTCAATACAGAGTCCTTTTCAGGGTCGTAGG GATGATCTTTAGATGGAATTTCTAAAATAGCAGGGAACGCATTGGTGAAGGAGTCCACTC

1500 1560 1620 1680 1740 1800 1860

V

H

A

TAGCTCTTT

V

R

L

R

P

S

L

T

F

E

E

K

H

A

D

I