Nucleic Acids Research, Vol. 18, No. 23 7137
©n) 1990 Oxford University Press
Nucleotide
sequence of RAD4 gene of Saccharomyces cerevisiae that can be propagated in Escherichia coli
without inactivation In Soon Choi, Jae Bum Kim and Sang Dia Park* Department of Zoology, College of Natural Sciences, Seoul National University, Seoul 151 -742, Korea Submitted October 31, 1990
The RAD4 gene of Saccharomyces cerevisiae, which is absolutely required for the nucleotide excision repair, was isolated from a yeast genomic library (1). The cloned RAD4 gene was able to propagate in Escherichia coli without any genetic change, to map into the RAD4 locus on chromosome V, and had the same restriction map as wild type yeast chromosomal DNA (2). Fleer et al. (3) independently isolated RAD4 gene by the gaprepair method. This cloned gene, however, was inactivated in E. coli. We have determined the nucleotide sequence of RAD4 gene. The sequence contains a 5' non-coding region of 92 bases followed by 2190 nucleotides showing a single continuous open reading frame starting with the initiation methionine at position 93. Sequence analysis showed that the RAED4 gene encodes for a putative protein of 730 amino acids with the molecular weight of 84.456, which is consistent with the transcript size of 2.3 kb and the Rad4 protein of 89 kDa (Kim et al., unpublished data). The TATAA sequence, starting at nucleotide position 14-18, is a candidate for TATA box. In the 3' non-coding region, the AATAAA sequence of polyadenylation signal is present at position 2321-2326. However, no significant homology between RAD4 gene and other genes was identified.
ACKNOWLEDGEMENT This work was supported by Korea Science and Engineering Foundation and Ministry of Education, the Republic of Korea to S.D. Park. REFERENCES 1. Yoon,J.K., Choi,I.S., Huh,S.O., Choe,S.Y. and Park S.D. (1985) Korean J. Genetics 7-3, 97-104. 2. Choi,I.S., Kim,J.B., Lee,K.N. and Park,S.D. (1990) Photochem. Photobiol. 52, 395-400. 3. Fleer,R., Nicolet,C.M., Pure,G.A. and Friedberg,E.C. (1987) Mol. Cell. Biol. 7, 1180-1192.
*
To whom correspondence should be addressed
EMBL accession
X55891
no.
X 102
V
AAGAT_CTAAGA A ACGAATATrDAAGAAAAATCAACAAA KS L K E T V I L K K N L N K W S W K R I K L S F L L K L V R K W K
204
306
A
K Q L H D S P S L X L L R N I I S A F K V A V N L N K E E N I E D Y 1 K Y A I T D E Y A L D E L N F K V L K D V P Q A I Q X N A P Y I I V
408 K G A R T L P N G G N V S R V S S I V K S H A G S L L I L L N D I T
510 N T E T A A L V L H S V N E LS
P Y L L S Y R R I L K E L I X S I V
612 G V W S TT R E L Q T Q I A S F A F L I N T T K E FP
1
SE
L E T T
714
L I Q R T S T F I X S C R X T N X R ES
P L I N F Q I N S A A E L F
816 91H 1020
G I D E V L G Y Q V G F E Y I R Q L A I H L R N T K N A T T K K S S A K I N S A E A Y K I V Y N W Q F C H S L D F W S R V L S F A C Q P E A K E N G S E S P L R Q L I Y P L V Q V T L G V I R L I P T P Q F F P
1122
L R F Y L I K S L I R L S Q N S G V F I P I Y P L L S E I L T S T A 1224
F T I A P X X S P N L A A F H F E H N I K C T Q A Y L N T I I Y Q E 1326
G L S E Q F V D L L G D Y F A L Y C I N I A F P E L V T P V I I S L 1428
R R Y I X T S T N V K L N 1530
I R L S T V V El L N Q N S T F I Q E K R
TCAGi S D V E F G P T N I S E V S R F L N D V A W N I T P L G S Y V A V Q
1632
R E V K E E X A R L
RS E S N E E Q DD
E R E T S E A X L L N S L E
1734
AELL S D D D N E D V E N V R R L T T Q Q D L I F C L
1836
A
E F N S I V Y K C V
1938 2040
GTrATrAAATrIT.AAAT.1TCAAcTCIVAAAD!T.CTCAaIAAATAGATATDATIDCrAACATAM36DAAAATrFTAGrACAADATATATDATITCr
2142 2244
Y C W X I S S N L N L G A N F A D N N S W N N S F A A Y G ATATT_ADCAcCADDrADAACACCAACCACTATA_TrAAT CA I F S A L L Q V V H Q P L Z
2346
A TWACDATeEADACT
1ATAAATE!CACCOAX1DACAACCAA
2431
©n) 1990 Oxford University Press
Nucleotide
sequence of RAD4 gene of Saccharomyces cerevisiae that can be propagated in Escherichia coli
without inactivation In Soon Choi, Jae Bum Kim and Sang Dia Park* Department of Zoology, College of Natural Sciences, Seoul National University, Seoul 151 -742, Korea Submitted October 31, 1990
The RAD4 gene of Saccharomyces cerevisiae, which is absolutely required for the nucleotide excision repair, was isolated from a yeast genomic library (1). The cloned RAD4 gene was able to propagate in Escherichia coli without any genetic change, to map into the RAD4 locus on chromosome V, and had the same restriction map as wild type yeast chromosomal DNA (2). Fleer et al. (3) independently isolated RAD4 gene by the gaprepair method. This cloned gene, however, was inactivated in E. coli. We have determined the nucleotide sequence of RAD4 gene. The sequence contains a 5' non-coding region of 92 bases followed by 2190 nucleotides showing a single continuous open reading frame starting with the initiation methionine at position 93. Sequence analysis showed that the RAED4 gene encodes for a putative protein of 730 amino acids with the molecular weight of 84.456, which is consistent with the transcript size of 2.3 kb and the Rad4 protein of 89 kDa (Kim et al., unpublished data). The TATAA sequence, starting at nucleotide position 14-18, is a candidate for TATA box. In the 3' non-coding region, the AATAAA sequence of polyadenylation signal is present at position 2321-2326. However, no significant homology between RAD4 gene and other genes was identified.
ACKNOWLEDGEMENT This work was supported by Korea Science and Engineering Foundation and Ministry of Education, the Republic of Korea to S.D. Park. REFERENCES 1. Yoon,J.K., Choi,I.S., Huh,S.O., Choe,S.Y. and Park S.D. (1985) Korean J. Genetics 7-3, 97-104. 2. Choi,I.S., Kim,J.B., Lee,K.N. and Park,S.D. (1990) Photochem. Photobiol. 52, 395-400. 3. Fleer,R., Nicolet,C.M., Pure,G.A. and Friedberg,E.C. (1987) Mol. Cell. Biol. 7, 1180-1192.
*
To whom correspondence should be addressed
EMBL accession
X55891
no.
X 102
V
AAGAT_CTAAGA A ACGAATATrDAAGAAAAATCAACAAA KS L K E T V I L K K N L N K W S W K R I K L S F L L K L V R K W K
204
306
A
K Q L H D S P S L X L L R N I I S A F K V A V N L N K E E N I E D Y 1 K Y A I T D E Y A L D E L N F K V L K D V P Q A I Q X N A P Y I I V
408 K G A R T L P N G G N V S R V S S I V K S H A G S L L I L L N D I T
510 N T E T A A L V L H S V N E LS
P Y L L S Y R R I L K E L I X S I V
612 G V W S TT R E L Q T Q I A S F A F L I N T T K E FP
1
SE
L E T T
714
L I Q R T S T F I X S C R X T N X R ES
P L I N F Q I N S A A E L F
816 91H 1020
G I D E V L G Y Q V G F E Y I R Q L A I H L R N T K N A T T K K S S A K I N S A E A Y K I V Y N W Q F C H S L D F W S R V L S F A C Q P E A K E N G S E S P L R Q L I Y P L V Q V T L G V I R L I P T P Q F F P
1122
L R F Y L I K S L I R L S Q N S G V F I P I Y P L L S E I L T S T A 1224
F T I A P X X S P N L A A F H F E H N I K C T Q A Y L N T I I Y Q E 1326
G L S E Q F V D L L G D Y F A L Y C I N I A F P E L V T P V I I S L 1428
R R Y I X T S T N V K L N 1530
I R L S T V V El L N Q N S T F I Q E K R
TCAGi S D V E F G P T N I S E V S R F L N D V A W N I T P L G S Y V A V Q
1632
R E V K E E X A R L
RS E S N E E Q DD
E R E T S E A X L L N S L E
1734
AELL S D D D N E D V E N V R R L T T Q Q D L I F C L
1836
A
E F N S I V Y K C V
1938 2040
GTrATrAAATrIT.AAAT.1TCAAcTCIVAAAD!T.CTCAaIAAATAGATATDATIDCrAACATAM36DAAAATrFTAGrACAADATATATDATITCr
2142 2244
Y C W X I S S N L N L G A N F A D N N S W N N S F A A Y G ATATT_ADCAcCADDrADAACACCAACCACTATA_TrAAT CA I F S A L L Q V V H Q P L Z
2346
A TWACDATeEADACT
1ATAAATE!CACCOAX1DACAACCAA
2431
