title: Implication of HpEts in gene regulatory networks responsible for specification of sea urchin skeletogenic primary mesenchyme cells.
abstract: The large micromeres of the 32-cell stage of sea urchin embryos are autonomously specified and differentiate into primary mesenchyme cells (PMCs), giving rise to the skeletogenic cells. We previously demonstrated that HpEts, an ets-related transcription factor, plays an essential role in the specification of PMCs in sea urchin embryos. In order to clarify the function of HpEts in the gene regulatory network involved in PMC specification, we analyzed the zygotic expression pattern and the cis-regulatory region of HpEts, and examined the activity of the HpEts protein as a transcription factor. Intron-based PCR reveals that zygotic expression of HpEts starts at the cleavage stage, and that the rate of transcription reaches maximum at the unhatched blastula stage. A series of progressive deletions of the fragments from -4.2 kbp to +1206 bp of the HpEts, which directs PMC-specific expression, caused a gradual decrease in the specificity, implying that coordination of several cis-regulatory elements regulates the expression in PMCs. A minimum cis-element required for the temporal expression is located within a 10 bp from -243 bp to -234 bp. The HpEts protein remains in the cytoplasm of entire embryonic cells in the cleavage stage. At the unhatched blastula stage, the HpEts protein translocates into the nucleus in presumptive PMCs. Transactivation assays demonstrate that the HpEts protein activates a promoter of Spicule Matrix Protein 50 (SM50), which is a target of HpEts, which binds to the regulatory region of SM50.
authors.affiliation: Department of Molecular and Cell Biology and Biochemistry, Brown University, Providence, RI 02912, USA.
authors.forename: Mamiko
authors.initials: M
authors.lastname: Yajima
authors.forename: Rieko
authors.initials: R
authors.lastname: Umeda
authors.forename: Takuya
authors.initials: T
authors.lastname: Fuchikami
authors.forename: Miho
authors.initials: M
authors.lastname: Kataoka
authors.forename: Naoaki
authors.initials: N
authors.lastname: Sakamoto
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Koji
authors.initials: K
authors.lastname: Akasaka
mesh_terms: D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D053263:Gene Regulatory Networks / Q000502:physiology*; D047330:Hemicentrotus / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D008969:Molecular Sequence Data; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D015053:Zygote / Q000502:physiology
chemical_list: D012333:RNA, Messenger; D014157:Transcription Factors