The RNA-binding protein Xp54nrb isolated from a Ca²⁺-dependent screen is expressed in neural structures during Xenopus laevis development

Abstract

ABSTRACT

Kcnip1 a Ca2+-dependent transcriptional repressor regulates the size of the neural plate in Xenopus

“…Stained embryos were then embedded in 3% low melting agarose for vibratome sectioning. For immunohistochemistry analysis, in vivo expression of GFP signals was performed on embryos fixed at stage 9 or stage 14 [13]. The proliferative cells were stained with anti-phospho-Histone 3 (Ser10) anti-body (1/1000e, Upstate) and revealed using an Alexa-555-coupled secondary anti-body (Fisher Scientific).…”

“…Protein-DNA complexes were resolved in 5% nondenaturing polyacrylamide gels and visualized by autoradiography. For Western blot analysis, control and microinjected embryos were lysed at stage 13 and 10 K supernatants equivalent to 1 embryo and were applied on SDS-PAGE as described [13]. The GFP and Kcnip1-GFP proteins were revealed by immunoblotting with rabbit anti-GFP polyclonal antibody (1/5000, TP401, Torrey Pines, Clinisciences) and goat peroxidase-conjugated anti-rabbit antibody (1/5000, Promega) was detected with the enhanced chemiluminescent kit (ECL, Amersham).…”

“…Through a differential screening between naive and Ca 2+ -stimulated ectoderms we have isolated genes encoding transcriptional regulators and RNA binding proteins downstream of the Ca 2+ signaling pathway [10][11][12][13]. Among the isolated genes we demonstrated that the arginine methyltransferase prmt1b acts as an early proneural gene that regulates zic3 transcription.…”

The calcium: an early signal that initiates the formation of the nervous system during embryogenesis

“…A total of about 400 clones were screened, and about 30 clones were found to selectively hybridize to the neural-subtracted probe and not to the epidermis-subtracted probe. Among these clones, xMLP encodes a MARCKS-like protein, a substrate for PKC (Zhao et al, 2001; Batut et al, 2003); xPRMT1b is the Xenopus homologue of the mammalian arginine methyltransferase PRMT1 gene (Batut et al, 2005); xId3 (Wilson and Mohun, 1995), encodes HLH protein that acts as dominant negative inhibitor of bHLH transcription factors; and Xp54nrb encodes a protein which exhibits the RRM domains characteristic of RNA binding proteins, and implicated in pre-mRNA splicing steps (Neant et al, 2011). The spatio-temporal expression pattern of these genes is restricted to neural territories and their expression is triggered following the inhibition of BMP signaling by noggin.…”

Ca2+ coding and decoding strategies for the specification of neural and renal precursor cells during development

“…Xmlp encodes a MARCKS-like protein, which is a substrate for protein kinase C (PKC) [26,28]; prmt1b is the Xenopus homologue of the mammalian arginine methyltransferase PRMT1 gene [29]; xId3 is an HLH factor that acts as dominant negative inhibitor of bHLH transcription factors [30]; and p54nrb encodes an RRM-domain protein characteristic of RNA-binding proteins and it has been implicated in pre-mRNA splicing steps [31]. The spatio-temporal expression pattern of these genes was shown to be restricted to the neural territories and their expression was triggered following an inhibition of BMP signalling by noggin.…”