Gene or accession: CG31049 Release: 3 cDNA or EST error Comments: As we published in 2000, multiple DOA proteins exist (we're up to 4), based upon data from western blots, confirmed with affinity-purified antibodies and use of mutants to confirm origin of the protein. We sequenced RE30665 in its entirety- this EST contains the N-terminal end of CG31049 and the C-terminal (Kinase domain) of Doa, CG1658, as we'd hoped, and thus it encodes one of the alternative Doa isoforms (of 105 kD). Given its placement in the gene, I suspect that CG33204 will also turn out to be a Doa isoform, but we've not done anything with that yet, nor do we have any experimental evidence to confirm it actually exists. Do you have any I'm missing in the database? Also, please note that a previous BLAST analysis we reported (Rabinow and Rabinow, 2001.5.23 Personal communication to FlyBase available from http://flybase.bio.indiana.edu/.bin/fbpcq.html?FBrf0135387 <up>FBrf0135387</up>) with sequence from one of the alternative cDNAs we'd obtained was partly misleading and should be deleted from the database if possible. Thank you very much. Best wishes, Lenny Rabinow Subject: Re: Doa/CG31049 correction >Dear Dr. Rabinow, > >It would be very helpful if you could send a FASTA file of RE30665 and the >encoded CDS to insure that we properly correct the Doa annotation. If you can >send it soon, I will try to include it in the upcoming annotation release. >Thanks for your help. > >Sincerely, > >Beverley Matthews >Curator, FlyBase-Harvard > Hello Beverly, .. Attached is a Word file (sorry for the improper format, we'll do better next time) of the complete DNA sequence of RE30665, supporting the fact that CG31049 is actually an alternately spliced (and initiated) isoform of Doa kinase (CG1658). Based on northern hybridizations and predicted protein size from the ORF, this message, reported in Yun et al 1994 as 4.0 kb FBrf0074875 Yun, B., R. Farkas, K. Lee, and L. Rabinow. 1994. The Doa locus encodes a member of a new protein kinase family, and is essential for eye and embryonic development in Drosophila melanogaster. Genes Dev. 8: 1160-1173 encodes the 105 kD cytoplasmic isoform of the kinase reported in Yun et al. 2000 FBrf0130184 Yun, B., K. Lee, R. Farkas, C. Hitte, and L. Rabinow. 2000. The LAMMER protein kinase encoded by the Doa locus of Drosophila is required in both somatic and germ-line cells, and is expressed as both nuclear and cytoplasmic isoforms throughout development. Genetics 156: 749-761. If you attribute this data, please cite A. Kpebe and L. Rabinow, unpublished (or in prep.) Thanks. As for another predicted gene in the introns of Doa, CG33204, we've no evidence either way regarding it. I'm willing to bet it's yet another DOA isoform. I note that the ESTs supporting this 'gene': RH25104; RH05514; RH28293; RH21287; RH06871; RH31228; RH70859 are all 5' sequences. I will order one or two of the above ESTs and we'll sequence from the 3' side (unless you can request the folks at the physical-end of BDGP to it first, since I'm sure they'll be quicker). Let me know if this is of any interest to you. Any insight into any of the above would be appreciated, With all the information available through Flybase it's easily possible I'm missing yet other annotations of interest to us on Doa. Thanks very much for your assistance. Best wishes, Lenny Rabinow
