IDENTIFICATION OF THE GENES-CODING FOR THE 2ND-LARGEST SUBUNITS OF RNA POLYMERASE-I AND POLYMERASE-III OF DROSOPHILA-MELANOGASTER

被引:19
作者
SEIFARTH, W
PETERSEN, G
KONTERMANN, R
RIVA, M
HUET, J
BAUTZ, EKF
机构
[1] UNIV HEIDELBERG,INST MOLEK GENET,NEUENHEIMER FELD 230,W-6900 HEIDELBERG,GERMANY
[2] CEA,SERV BIOCHIM & GENET MOLEC,F-91191 GIF SUR YVETTE,FRANCE
来源
MOLECULAR & GENERAL GENETICS | 1991年 / 228卷 / 03期
关键词
D O I
10.1007/BF00260636
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have isolated cDNA and genomic clones of Drosophila melanogaster by cross-hybridization with a 658 bp fragment of the yeast gene coding for the second-largest subunit of RNA polymerase III (RET1). Determination of the sequence by comparison of genomic and cDNA regions reveals an ORF of 3405 nucleotides which is interrupted in the genomic sequence by an intron of 48 bp. The deduced polypeptide consists of 1135 amino acids with a calculated molecular weight of 128 kDa. The protein sequence shows the same conserved regions of homology as those observed for all the second-largest subunits of RNA polymerases cloned so far. The gene (DmRP128) obviously codes for a second-largest subunit of an RNA polymerase which is different from DmRP140 and DmRP135. We have purified three distinct RNA polymerase activities from D. melanogaster. By using specific RNA polymerase inhibitors in enzyme assays and by comparing their subunit composition we were able to distinguish between RNA polymerase I, II, and III. RNA polymerase preparations of D. melanogaster were blotted and the second-largest subunits were identified with antibodies raised against polypeptides expressed from DmRP128 and DmRP135. Anti-DmRP135 antibodies react strongly with the second-largest subunit of RNA polymerase I but do not react with the respective subunits of RNA polymerase II and III. The second-largest subunit of RNA polymerase III is only recognized by anti-DmRP128. Previously, we have claimed that DmRP135 codes for the second-largest subunit of RNA polymerase III. Based on the new biochemical data reported here we show that DmRP135 codes instead for the second-largest subunit of RNA polymerase I and that DmRP128 corresponds to the equivalent subunit of RNA polymerase III.
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页码:424 / 432
页数:9
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