©~1990 Oxford University Press
4256 Nucleic Acids Research, Vol. 18, No. 14
Nucleotide and derived amino-acid sequence of the major intrinsic protein of rat eye-lens
Nicholas A.Kent and Alan Shiels School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK Submitted June 11, 1990
EMBL accession no. X53052
We have isolated and sequenced a cDNA coding for the major intrinsic protein (MIP) of rat eye-lens fibre cell-membranes. The MIP clone was isolated from a rat lens cDNA library (XgtlI 1) using an oligonucleotide probe derived from the bovine MIP sequence (1). Both strands of the cDNA were sequenced by the dideoxy chain termination method using Sequenase (USB) and variations between the rat and bovine MIP sequences are indicated. The rat MIP cDNA contains a 786 bp open reading frame followed by a 3' untranslated region which includes a putative polyadenylation signai (underlined). The initiai 37 Nterminal amino-acids deduced from the rat cDNA sequence are identical to those of rat MIP determined by Edman degradation (2) and overall rat and bovine MIP are -94 % identical at the amino-acid level. Amino-acids enclosed in boxes are conserved Bcv Rat Rat
Bcv Bov Rat Rat Dcv Bov Rat Rat Dcv Dcv Rat Rat
Bov Bov Rat Rat Boy Bov Rat Rat Boy Bov Rat Rat
Bcv Bov Rat Rat
Boy Bov Rat Rat Bov Bov Rat
ATO TOG N
W
G
A
between members of an evolutionarily diverse family of transmembrane 'channel' proteins (3).
ACKNOWLEDGEMENT
Supported by the Wellcome Trust. REFERENCES 1. Gorin,M.B., Yancey,S.B., Cline,J., Revel,j.-P. and Horwitz,J. (1984) Cell
39, 49-59. 2. Nicholson,BJ., Takemoto,L.J., Hunkapiller,M.W., Hood,L.E. and Reve1,j.P. (1983) Cell 32, 967-978. 3. Rao,Y., Jan,L.Y. and Jan,Y.N. (1990) Nature 345, 163-167.
C
C
GAA CTT CBS TCT GCC TCC TTT TGG AGO 0CC ATA TTT GCT GAG TTC L R S E A F S A I F A W R 9E F C
C G A 66a TTIT GiCT A.X' (Tr- rwc TAT 1311 rTC TTT UP6 CTG GGA TCC f F A Y V F L L.fG 6, s [D .s A
C T 6 T a C T 6i TCA CTO CST TG6 6CC CCT GGA CCC CTA CAT GTC CTG CAA GTG OGCT TTG GCC T'TT 0G1: C6 6CTO 6CC ACA N R 6 S L A P F P L H V L 0 V A L A A L- A I, A CAC ATC A6T GOA 6CC CAT GTC AAC CCI 6CR) H I A P A IN
ElV
A TC C C 6CA DCC CAG CTT CT6 66A OCT 9 A A L L A V 6 T CC T CTO CAT GCT 666 OTA AGC 616 H A V L a S V P C GAG AGG CB6 AAT GGC C6C ATG 6 R E R N M R L A C T C DCG 666 ATG AAT CCC 6CC C6C 6 A I AMRI
9]
A
CA
C
T
C
6
6TG GCT GOA V
6
A
0 C CAG ACT GTG GOT G T V a A 6 C T CA C rTC ACT TTT GCC TT'C CTT TGT 6CC TCC CAR ATSG TCC rro CTC CI3T OiCT TTC TGC TAT ATA 9 6 Ii S V I F F L V L] L L A F S, C r, I H C C C C. T T T GCG GCT GTG CT6 TAC AGT GITC ACC CCA C(':A ;rCT OTC CI3A 13GA AACr CI.A RCA CTC AAC AC6 9 1 6 6 A A V S V P A V Y P 1H L A L N T
1rITG
L
V
TA
BBC CAB 6CC ACC ACT GTG GAG ATC T'TC CTG D
a
A
T
C
T I
F
V
I
F
AC6G CTC CAG
L
a
T'IC F-
13'6 V
C
CTC TGC L C
ATIC I
TTT F
A
GCC ACG. A
TI
C
C TAT V
GAC
TCC TTT GCT CCT 9 S F A T
C
CTIC ACC AGA AAC ]'TC AIJC AAiC CALC lAGT c- rAc :(L 16 r6rG
L-
T
R
N
F
S T
H
H
W
V
V
W
T
BGC 666 66C CTG G6T ABC CT6 C'TC TAC 6AC 'ITT CTC C IC TIf C CCA COG CTC AAG AiT 6 6 L S L F L G6 L F L D LP fl K, 9 A
6 6 A Tr. AGA CCC A6T SAC TCC AAT 6GA CAB CCA GAG GGT ACA GOG6 AA CCT 6 6 F P 0 6 S F, R S N E T () E E V
T
GT6 V
SjI'T rlCT GAG- ACiA CTO FC1' T S V
F.
C GAA CTG AAG ACT CAR 1I L T VF E
L
L
V
T ATC I
66 T C CCA AI'C AlT C 6 1 I V TT
C,TC AA6 OIGA 6CC L
1'.
180 60
270 90
360 120 450 150
T
T
G6CC' TCT OTO CCC CTG OC~T GrT GM C TTC TUC CTC. ACC CIA T 3rCAi CAC CIE 'TIT (irir ATO IAT TAC ACT G6C S 6 V A L A V S F L L T r6 H- L F C, M Y Y T 6, C T C Ai L A I
90 30
a
A s
qaAr.tc:caqc:cgq.tagagg~gg TAA *layJ:ycagaacgccgc:tgggtt WCM CTR, K )
540
1830 630
21) 720
24')
815 26
qtcjtt taggagctttgggttacaggagggctagccgccccggataari~%;Aagac.qtgggagggatgtcttt-Ltttl-icttl ttttttgtgttttttttttgcct
tcggaggcaccgcctgaga.A:aggcttggag!gatatggattttattt___a_gt_
934tctttctttgatc 3
4256 Nucleic Acids Research, Vol. 18, No. 14
Nucleotide and derived amino-acid sequence of the major intrinsic protein of rat eye-lens
Nicholas A.Kent and Alan Shiels School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK Submitted June 11, 1990
EMBL accession no. X53052
We have isolated and sequenced a cDNA coding for the major intrinsic protein (MIP) of rat eye-lens fibre cell-membranes. The MIP clone was isolated from a rat lens cDNA library (XgtlI 1) using an oligonucleotide probe derived from the bovine MIP sequence (1). Both strands of the cDNA were sequenced by the dideoxy chain termination method using Sequenase (USB) and variations between the rat and bovine MIP sequences are indicated. The rat MIP cDNA contains a 786 bp open reading frame followed by a 3' untranslated region which includes a putative polyadenylation signai (underlined). The initiai 37 Nterminal amino-acids deduced from the rat cDNA sequence are identical to those of rat MIP determined by Edman degradation (2) and overall rat and bovine MIP are -94 % identical at the amino-acid level. Amino-acids enclosed in boxes are conserved Bcv Rat Rat
Bcv Bov Rat Rat Dcv Bov Rat Rat Dcv Dcv Rat Rat
Bov Bov Rat Rat Boy Bov Rat Rat Boy Bov Rat Rat
Bcv Bov Rat Rat
Boy Bov Rat Rat Bov Bov Rat
ATO TOG N
W
G
A
between members of an evolutionarily diverse family of transmembrane 'channel' proteins (3).
ACKNOWLEDGEMENT
Supported by the Wellcome Trust. REFERENCES 1. Gorin,M.B., Yancey,S.B., Cline,J., Revel,j.-P. and Horwitz,J. (1984) Cell
39, 49-59. 2. Nicholson,BJ., Takemoto,L.J., Hunkapiller,M.W., Hood,L.E. and Reve1,j.P. (1983) Cell 32, 967-978. 3. Rao,Y., Jan,L.Y. and Jan,Y.N. (1990) Nature 345, 163-167.
C
C
GAA CTT CBS TCT GCC TCC TTT TGG AGO 0CC ATA TTT GCT GAG TTC L R S E A F S A I F A W R 9E F C
C G A 66a TTIT GiCT A.X' (Tr- rwc TAT 1311 rTC TTT UP6 CTG GGA TCC f F A Y V F L L.fG 6, s [D .s A
C T 6 T a C T 6i TCA CTO CST TG6 6CC CCT GGA CCC CTA CAT GTC CTG CAA GTG OGCT TTG GCC T'TT 0G1: C6 6CTO 6CC ACA N R 6 S L A P F P L H V L 0 V A L A A L- A I, A CAC ATC A6T GOA 6CC CAT GTC AAC CCI 6CR) H I A P A IN
ElV
A TC C C 6CA DCC CAG CTT CT6 66A OCT 9 A A L L A V 6 T CC T CTO CAT GCT 666 OTA AGC 616 H A V L a S V P C GAG AGG CB6 AAT GGC C6C ATG 6 R E R N M R L A C T C DCG 666 ATG AAT CCC 6CC C6C 6 A I AMRI
9]
A
CA
C
T
C
6
6TG GCT GOA V
6
A
0 C CAG ACT GTG GOT G T V a A 6 C T CA C rTC ACT TTT GCC TT'C CTT TGT 6CC TCC CAR ATSG TCC rro CTC CI3T OiCT TTC TGC TAT ATA 9 6 Ii S V I F F L V L] L L A F S, C r, I H C C C C. T T T GCG GCT GTG CT6 TAC AGT GITC ACC CCA C(':A ;rCT OTC CI3A 13GA AACr CI.A RCA CTC AAC AC6 9 1 6 6 A A V S V P A V Y P 1H L A L N T
1rITG
L
V
TA
BBC CAB 6CC ACC ACT GTG GAG ATC T'TC CTG D
a
A
T
C
T I
F
V
I
F
AC6G CTC CAG
L
a
T'IC F-
13'6 V
C
CTC TGC L C
ATIC I
TTT F
A
GCC ACG. A
TI
C
C TAT V
GAC
TCC TTT GCT CCT 9 S F A T
C
CTIC ACC AGA AAC ]'TC AIJC AAiC CALC lAGT c- rAc :(L 16 r6rG
L-
T
R
N
F
S T
H
H
W
V
V
W
T
BGC 666 66C CTG G6T ABC CT6 C'TC TAC 6AC 'ITT CTC C IC TIf C CCA COG CTC AAG AiT 6 6 L S L F L G6 L F L D LP fl K, 9 A
6 6 A Tr. AGA CCC A6T SAC TCC AAT 6GA CAB CCA GAG GGT ACA GOG6 AA CCT 6 6 F P 0 6 S F, R S N E T () E E V
T
GT6 V
SjI'T rlCT GAG- ACiA CTO FC1' T S V
F.
C GAA CTG AAG ACT CAR 1I L T VF E
L
L
V
T ATC I
66 T C CCA AI'C AlT C 6 1 I V TT
C,TC AA6 OIGA 6CC L
1'.
180 60
270 90
360 120 450 150
T
T
G6CC' TCT OTO CCC CTG OC~T GrT GM C TTC TUC CTC. ACC CIA T 3rCAi CAC CIE 'TIT (irir ATO IAT TAC ACT G6C S 6 V A L A V S F L L T r6 H- L F C, M Y Y T 6, C T C Ai L A I
90 30
a
A s
qaAr.tc:caqc:cgq.tagagg~gg TAA *layJ:ycagaacgccgc:tgggtt WCM CTR, K )
540
1830 630
21) 720
24')
815 26
qtcjtt taggagctttgggttacaggagggctagccgccccggataari~%;Aagac.qtgggagggatgtcttt-Ltttl-icttl ttttttgtgttttttttttgcct
tcggaggcaccgcctgaga.A:aggcttggag!gatatggattttattt___a_gt_
934tctttctttgatc 3
