Different modes of regulation of transcription and pre-mRNA processing of the structurally juxtaposed homologs, Rnf33 and Rnf35, in eggs and in pre-implantation embryos.

Molecular events involved in gene expression in unfertilized eggs and pre-implantation embryos are beginning to be understood. In this work, we investigated the transcription and processing of two structurally juxtaposed mouse RING finger protein genes, Rnf33 and Rnf35. Transcripts of these genes are detected only in eggs and in pre-implantation embryos. Both genes are intronless except for a solitary intron in the 5'-untranslated region. Here, we showed by rapid amplification of cDNA ends (RACE) and reverse transcription experiments that Rnf35 transcription uses a single promoter and a terminating site. On the other hand, Rnf33 is transcribed using multiple promoters. At the four-cell stage, however, Rnf33 mRNA with a single transcription start site derived from the proximal promoter is detected, indicating that it is the major promoter. Sequences upstream of the Rnf35 and the major Rnf33 transcription start sites carry no TATA boxes but a putative transcription initiator (Inr) element is discernible in each case. The processing of the 3'-end of the Rnf33 mRNA is also in disarray with multiple 3'-ends, an event that may be related to the absence of the AAUAAA element and the utilization of AAUAAA-like proxies. The multiplicity of the 3'-untranslated region is partially amended at the four-cell stage when only two major 3'-ends are in use. This work demonstrates that expression of some maternal and early zygotic genes may be opportunistic until a stringent transcriptional regulation mechanism is imposed.