Alternative transcripts of a polyhomeotic gene homolog are expressed in distinct regions of somites during segmentation of zebrafish embryos.
Here we describe isolation and characterization of two zebrafish cDNAs, designated ph2alpha and ph2beta, which were identified as structural homologs of the Drosophila polyhomeotic, mouse Mph2, and human HPH2 genes, collectively termed the Polycomb group. The alpha and beta transcripts shared a 1.9-kb sequence at their 3'-termini. Alpha had an additional 1.6-kb sequence extending toward its 5'-terminus. Only a short 0.1-kb segment was unique to beta. Sequencing of a genomic clone corresponding to the two cDNAs indicated that the mRNAs were transcribed from a single gene locus by alternative promoters. Northern blots revealed expression of alpha transcripts during the segmentation period, while beta expression occurred at all developmental stages examined. Whole-mount in situ hybridizations with an alpha-specific probe and a probe recognizing both transcripts revealed distinct spatio-temporal expression patterns along developing somites. Alpha transcripts were detected initially at the 7-8 somite stage; beta transcripts appeared in the first somites. As segmentation proceeded, alpha and beta expression shifted position toward the tailbud in parallel with the formation of each somite. Within individual somites, the signal corresponding to alpha was strongest at the posterior border and weakest in the anterior region. Conversely, that corresponding to beta was strongest at the anterior border and weakest in the posterior region. The data support the idea that Ph2alpha and Ph2beta are involved in spatio-temporal generation of somites as well as in specification of antero-posterior regional differences within individual somites.
Kawamura
Akinori
A
Department of Biology, School of Education, Waseda University, 1-6-1 Nishi-Waseda, Shinjuku, Tokyo 169-8050, Japan.
Yamada
Kouji
K
Fujimori
Ken'ichi
K
Higashinakagawa
Toru
T
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D019521:Body Patterning; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D029721:Drosophila Proteins; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005786:Gene Expression Regulation; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009698:Nucleoproteins / Q000096:biosynthesis / Q000235:genetics*; D010802:Phylogeny; D063150:Polycomb Repressive Complex 1; D011401:Promoter Regions, Genetic; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D016415:Sequence Alignment; D019170:Somites / Q000378:metabolism*; D024363:Transcription Initiation Site; D014158:Transcription, Genetic; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D029961:Zebrafish Proteins / Q000096:biosynthesis / Q000235:genetics*
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D009698:Nucleoproteins; D020033:Protein Isoforms; D012333:RNA, Messenger; D029961:Zebrafish Proteins; C072820:ph-d protein, Drosophila; C449648:phc2a protein, zebrafish; D063150:Polycomb Repressive Complex 1