Maragh Samantha, Miller Ronald A, Bessling Seneca L, Wang Guangliang, Hook Paul W, McCallion Andrew S
Biochemical Science Division, National Institute of Standards and Technology, Gaithersburg, Maryland, United States of America; McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.
McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.
PLoS One. 2014 Aug 29;9(8):e105460. doi: 10.1371/journal.pone.0105460. eCollection 2014.
We recently demonstrated that the gene encoding the RNA binding motif protein 24 (RBM24) is expressed during mouse cardiogenesis, and determined the developmental requirement for its zebrafish homologs Rbm24a and Rbm24b during cardiac development. We demonstrate here that both Rbm24a and Rbm24b are also required for normal somite and craniofacial development. Diminution of rbm24a or rbm24b gene products by morpholino knockdown resulted in significant disruption of somite formation. Detailed in situ hybridization-based analyses of a spectrum of somitogenesis-associated transcripts revealed reduced expression of the cyclic muscle pattering genes dlc and dld encoding Notch ligands, as well as their respective target genes her7, her1. By contrast expression of the Notch receptors notch1a and notch3 appears unchanged. Some RBM-family members have been implicated in pre-mRNA processing. Analysis of affected Notch-pathway mRNAs in rbm24a and rbm24b morpholino-injected embryos revealed aberrant transcript fragments of dlc and dld, but not her1 or her7, suggesting the reduction in transcription levels of Notch pathway components may result from aberrant processing of its ligands. These data imply a previously unknown requirement for Rbm24a and Rbm24b in somite and craniofacial development. Although we anticipate the influence of disrupting RBM24 homologs likely extends beyond the Notch pathway, our results suggest their perturbation may directly, or indirectly, compromise post-transcriptional processing, exemplified by imprecise processing of dlc and dld.
我们最近证明,编码RNA结合基序蛋白24(RBM24)的基因在小鼠心脏发生过程中表达,并确定了其斑马鱼同源物Rbm24a和Rbm24b在心脏发育过程中的发育需求。我们在此证明,Rbm24a和Rbm24b对于正常的体节和颅面发育也是必需的。通过吗啉代敲低减少rbm24a或rbm24b基因产物会导致体节形成的显著破坏。基于原位杂交对一系列与体节发生相关的转录本进行详细分析,结果显示编码Notch配体的环状肌肉模式基因dlc和dld及其各自的靶基因her7、her1的表达降低。相比之下,Notch受体notch1a和notch3的表达似乎没有变化。一些RBM家族成员与前体mRNA加工有关。对注射了rbm24a和rbm24b吗啉代的胚胎中受影响的Notch信号通路mRNA进行分析,发现dlc和dld有异常转录片段,但her1或her7没有,这表明Notch信号通路成分转录水平的降低可能是由于其配体的异常加工所致。这些数据表明Rbm24a和Rbm24b在体节和颅面发育中存在以前未知的需求。虽然我们预计破坏RBM24同源物的影响可能超出Notch信号通路,但我们的结果表明,它们的扰动可能直接或间接损害转录后加工,以dlc和dld的不精确加工为例。