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2942 Nucleic Acids Research, 1993, Vol. 21, No. 12 Complete nucleotide sequence of an archaeal (Pyrococcus encoding a homolog of eukaryotic transcription factor IIB (TFIIB) woeset) gene Roberta Creti, Paola Londei and Piero Cammarano* Istituto Pasteur-Fondazione Cenci Bolognetti, Universita di Roma 1, Dipt. Biopatologia Umana, Sez. Biologia Cellulare, Policlinico Umberto 1, Viale Regina Elena 324, 00161, Rome, Italy EMBL accession no. X70668 Received May 7, 1993; Accepted May 13, 1993 We have previously reported the sequence of the streptomycin (str) operon-equivalent of the archaeum Pyrococcus woesei (1). Subsequently, Ouzounis and Sander found that a partial ORF located downstream (and on the complementary strand) of the P.woesei str-operon-equivalent cluster, encodes the C-terminal 152 amino acids of a polypeptide that shares significant similarity with the eukaryotic transcription factor TFIB (2). We have now sequenced the P. woesei DNA region encompassing the missing (N-terminal) portion of the TFIIB gene and the upstream 296 nucleotides (EMBL Data Bank accession number X70668). The archeal gene is preceeded (at -73) by a consensus promoter. The structural similarities between the P. woesei and the eukaryotic TFIIB are shown in Figure 1. The archaeal sequence (300 amino acids) is 27.5% -33.8% identical with that of eukarya and exhibits unique structural motifs characteristic of eukaryotic TFII (4-6) such as (i) an imperfect amino acid repeat (encompassing residues 136-182 and 232-278) with 38.3% identity and 63.8% similarity between the N-terminal and the C-terminal repeats (ii) an intervening sequence characteristically rich (32%) in basic amino acids between repeats (iii) Zn(ll)-finger motif (Cys-X2-Cys-XI5-Cys-X2-Cys) located in close proximity to the N-terminus and (iv) a predicted amphipatic ca-helix sitite at the end of the N-terminal repeat (residues 167-184, P. woesei numbering). On the whole, the distinguishing structural motifs of TFlIB appear to predate the phylogenetic divergence of archaea and eukarya and have been stringently conserved upon ensuing evolution. ACKNOWLEDGEMENT R.Creti is recipient of a fellowship from the Fondazione A. Buzzati-Traverso. REFERENCES 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. Creti,R. et al. (1991) J. Mol. Evol. 33, 332-342. Ouzounis,C. and Sander,C. (1992) Cell 71, 189-190. Sharp,P.A. (1991) Nature 351, 16-18. Brown J.W. et al. (1989) CRC Crit. Rev. Microbiol. 16, 287-338. Malik S. et al. (1991) Proc. Natl. Acad. Sci. USA 88, 9553-9557. Pinto,I. et al. (1992) Cell 68, 977-988. Ha,I. et al. (1991) Nature 352, 689-695. Wampler,S.L. and Kadonaga,J.T. (1992) Genes and Develop. 6,1542-1552. Hisatake,K. et al. (1991) Nucleic Acids Res. 19, 6693. Tsuboi,A. et al. (1992) Nucleic Acids Res. 20, 3250. r I E X C l S I E -0-PCE-SELMIYIWEIICIVCltCYVIEENI INNE SU&-KMLEILLDCTE TULMEEA I ITSIISAMIRIVLrTCPEClVPKVEWlSEOCAL WCLC6LV1501SL TTlT_K__LWAClK-Es-D-EVWAICIllLCUL9LIUKA STlC ----_US AFKEISIwLWTI_1 _m I A_SPIEDYRM lUallVVGICSEVUCLMW C"Vl-So"TSElCrGtG @EE NKAAASE-LUMLTNI-lrTCSF xOL I AUSRID-LK-TCAN-DLLVEYLMCECOLVVMRSEC 11nNPIIK 640 1 N"SASTAL_TV-CANILAIVEDYL6ICttCOLVTIMSF M-ATOLW6STWISOTNI-W-TASKA MMISf KVEIT TLPIVST SF 64.1 PASTSIL IIALP-TCPOHAIALVEDYLIUlC Et6LVVMWVSTFSlllAT-Tla_ UWaLSTHI-IMT ASF Of.WUNSSM FICETTNIlY0I9 FW se See 153 US 1S2 1f9 140 1tZ UO 3Y06Nf nLISIASIL6tAETFEIHf61TTLNIIIL6GKSE06FLKlOTOllIlFCSNLGLnSAEYAOtE lllT TAWSLNLLUalPITWtlTVT B 3LYIACAlE6TFKEICAVSKISIOICFl-TUtA-LETSVLTIFTAfICftTMIAKAtAIVISlAAAlysASEIlIMFAWT 6 am 140 2$3 XL. 14 26T-4TLITOM 6t4 141 N.M 141 LF EQXSUWMANUIASACLYIACll;MTFKEICAV5KINUR190CFK6FL-ILIAC----S9LITTGO 10_ FllStFCt5LC - THIAFXAVELOLVM tlNlYAAAIVMSCA MTUELA6VA&VT 264 FC¢S"LTIIYLl"lXC AiT L264LYlSCLfIACAYT*IClt¢FKLILA Pvm 282 Sme 309 EYLV-EKLKIKVPIA t30 ILYBNtNM.V6GLIAN6WSLSNLP6VE 345 26 I6SSYIOJIWAKLFPEMfKFTTPI-6ULP9--- 315 265 Xto IIQOSLIYPAPSLFPA6FKF6TPV-LPUL- 316 mm 640 N. I25 LIIPRAPLF MFKFDTV-6ICLPL-285 Z105OtLIYP6APOLFPTDFKFOTPV-OKLPL- 316 316 FM sme 71.3s Sm 33.6 36.1 XtL 32.7 36.6 64t 32.7 36.0 79.0 79.4 94.3 Nsa 32.7 36.0 79.4 94.3 99.7 Figure 1. Alignment of the P.woesei TFIB sequence with those from Saccharomyces cerevisiae (Sce) (5), Drosophila melanogaster (Dme) (7), Xenopus laevis (Xa) (8), Homo sapiens (Hsa) (6) and rat (9). Asterisks identify the Zn finger motifs. Arrows indicate the N-terminal and the C-terminal elements of the direct repeat; plus symbols identify the putative amphipatic a-helix of the P.woesei TEIB sequence. Pairwise identities (percent) between all sequences are shown in a matrix form. * To whom correspondence should be addressed