Xenopus laevis - Europe PMC

k.) 1990 Oxford University Press

Nucleic Acids Research, Vol. 18, No. 15 4603

Isolation of two different c-ets-2 proto-oncogenes in Xenopus laevis C.-M.Wolff, P.Stiegler, M.Baltzinger, D.Meyer, J.Ghysdaell, D.SteheIin1, N.Befort and P.Remy Laboratoire de Biochimie, Institut de Biologie Moleculaire et Cellulaire du CNRS, 15 rue Rene Descartes, 67084 Strasbourg Cedex and 'INSERM U186/CNRS URA1160, Institut Pasteur, 1 rue Calmette, 59019 Lille, France EMBL accession nos. X51826, X52635

Submitted June 27, 1990

The c-ets proto-oncogene has been identified in two forms, cets-] and c-ets-2, in several species. By screening a Xenopus laevis cDNA library prepared from stages 1/H oocyte mRNAs with a chicken c-ets-2 cDNA probe, two different c-ets-2 cDNAs have been identified. These cDNAs of 1.1 kb and 1.3 kb, named Xl-ets-2a and Xl-ets-2b respectively, share 91.3% similarity. They most likely arise from two different Xl-ets-2 genes rather than from different alleles. When compared with c-ets-2 genes from humans, mouse and chicken, both Xenopus genes show similarities ranging from 66.5% to 72.7%. The two Xenopus c-ets-2 cDNA partial sequences encode polypeptides of 352 amino acids which share 95.2% identical residues; if conservative amino acid changes are considered, the

90

1

Xl-ets-2b Xl-ets-2a 124




GAA TTC TCC TTG GAG C E L S F E Q L S F E 91 GCA CCT GAC TTC GTT T T V F A P D V F D P A 181 CAT TCT AAC CGG GAC A S N R D H D S N Q H 271 AAT TAC CCT AAA AAT N N

Y Y

P P

K K

N N

361


CCC AGC TCT TGC CTT P P

S S

S S

C C

L L

451 Xl-ets-2b Xl-ets-2a 274


*

sequence similarity is 97.7%. The Xl-ets-2 polypeptides are highly conserved with 68 to 70% of their amino acids identical to those of the human, mouse, and chicken ets-2 proteins. The most conserved region maps in the C-terminus of the protein and contains both the DNA binding site and the nuclear translocation signal. Northern blot analysis revealed that the Xl-ets-2 genes are transcribed during oogenesis and embryonic development into a major 3.2 kb RNA and a minor 1.7 kb transcript. The same transcripts were also detected in a variety of adult tissues (brain, eye, liver, stomach, spleen, heart, kidney, muscle, ovary and testis).

CTA AAT TCT GGC AAA C G K G L N S K G S L N 541 AGC CAA TCT TCT CTT G S S L S Q S L S S Q

AAC GTG AAT TTT CAG AAG TTT CTC ATG AAT GGA CAC GAG CTA TGC AGC CTC GGG AAG GAG AGG TTT TTG GCT CTG G A K F L M N G H E L C S L G K E R F L A L Q F N V N F L M N G H E L C S K L G K E R F L A L Q F N V N 180 GGG GAC ATA CTT TGG GAA CAT CTG GAA GAA ATG ATG AAA GAG TAT CAA GAA AAG GCC CAG GAG CCA TAT ATT GAC G C A C A C G Y H L E E M M K E E K A Q E P Y I D E Q W L I D G L E E M M K E H Q H E K A Q E P Y I D E W L I D G 270 TCA CTA AAC CAG TGG ATG AAT GCA GAT TCC TTA AAT TTC ACT GCT GAT CCG TTG CAG TGT GGG GCA CAA GTA CAC C A T A T C A P A N D S L N F T A D L Q C G A Q V H M W S Q N L C S L N F T T D P L Q G A Q V D H A N M W H N I S

360 GGA ATG TAT AAT GAT ATG TGC TCA GTG CCC ACT GGT CAG ACT CTC CTC AAC CCC AAA CAA GAG TTC CAA CAA TAC T A C A T Y S C V P T G Q T L L N P K Q E F Q Q M D N Y M G P T L L Q K Q E F Y G T Q P Q V S C M D Q N F M G 450 AAA TCC AGA GCA GTA AAC TAC CCG CCC GCC AGC CAG GAC TTT GCA AGG AGC CAT ATG AAC GTG TTG CTC AAC TCT T A A T CA C T G A S D F A R S H M N V L L N S P P Y N V A Q R S K F A R Q S N M N A L L N S S A P S Y D N V A R S K 540 TTG AGA GAC TAT GAC TCA GGA GAC AGC GGC ACA GAA AGC TTC GAG AGC ACC GAA TCA CTA CTG CAT TCT TGG ACG G T T A T G T S T E S L L H S W E T F S E T G S D G S D Y D R L S S S S T E F E T E L L Q W T G S D G S D Y D R L 630 GTC GAT ATG CAA CGA GTT CCT TCA TAT GAC AGC TTT GAG GAA GAT GGC AAT CAA ACG TTG TGT CTA AAT AAG CAA C GC G G A C G C T C S F E E G N Q T L C L N K Q D S Y M P V D R Q D V C S E E D G A L L N K P F Q G S D Y P V R Q D V M

To whom correspondence should be addressed

4604 Nucleic Acids Research, Vol. 18, No. 15 Xl-ets-2b Xl-ets-2a 334




720 631 CCA ATG TCC TTC AAA GAT TAT ATC CAG GAC AGG TGT GAG CCA GCA GAA CTA GGC AAA CCT GTT ATT CCC GCA TCT ATA CTT GCT GGC TTC G G A T A A C T R C L K P V I A S I L A G F Y I Q D P A E G P F K D E P M S V I Y R E L G K P A S I L A G F P F K D I Q D C E P A P M S 810 721 ACA GGC AGT GGA CCG ATC CAA TTG TGG CAG TTC TTG TTG GAA CTT TTA ACG GAT AAG TCA TGT CAG TCC TTT ATC AGC TGG ACT GGG GAT G C A A G C A Q Q S C S G K S F I S W T G D T G P I L W F L L E L L T D Q S F I S W T G D G S G P I Q L W Q F L L E L T D K S C Q T L 900 811 GGA TGG GAG TTC AAG CTG GCT GAC CCA GAT GAG GTG GCC CGG CGG TGG GGT AAA AGG AAA AAC AAA CCT AAA ATG AAC TAC GAG AAG CTA C C A T G W E F K L E A D P D V A R R W G K R K N K P K M N Y E K L W E F K L T E G D P D E V A R R W G K R K N K P K M N Y K L

901 990 AGT CGA GGA CTA AGG TAT TAT TAT GAC AAG AAC ATC ATC CAC AAG ACA TCA GGG AAG AGA TAC GTG TAC CGA TTT GTT TGT GAC CTT CAC C C C C T G G L R Y Y R G Y D K N I S S I H K T G K R Y V Y R F V C D L H R G L R Y Y K S Y D N I K T S R I H G K F L Y V Y R V C D H

991 Xl-ets-2b

Xl-ets-2a 454 Xl-ets-2b Xl-ets-2a Xl-ets-2b

1087

AAC TTA TTG GGC TAC ACG CCG GAT GAA TTG CAC GCA ATG CTT GGA GTA CAG CCC GAC ACA GAC GAA TGAGCTGCCGGAGTGTGAACTAAATCATTGG C C C C G T A T G N L G Q * L Y T P D E L H A M L G V P D T D E Q * N L L Y T G P D E L H A D M L G V P D T E

1088 1206 TTTGAAGAGCACATTATTGTACTGTGAAAAGGATCGTTTAGTGGGCTGTCCAAGATGGCCGAACAGTGCATGACTTCAAAAGACTGGAAACATCAAATCCGTGACATGAATTGGCTCTT C C A 3'-end 1285 TGCTCTGTTTTCTTAGATCGGAGTAAAGATCTGATGGATGGCGCTTATTGTTATACAGTTGCTCACTTGAAAAAAAAAA