Biomedical subjects

Scott Rankin

Publications and source records attributed to Scott Rankin.

5 recordsLinked to original sources

PubMed2004

Smad2 and Smad3 coordinately regulate craniofacial and endodermal development.

Ligands of the transforming growth factor-beta (TGF-beta) superfamily are involved in numerous developmental and disease processes. TGF-beta, activins, and nodal ligands operate through the highly homologous Smad2 and Smad3 intracellular mediators. Smad2 mutants exhibit early embryonic lethality, while Smad3 mutants are viable, but show a plethora of postnatal phenotypes, including immune dysfunction and skeletal abnormalities. Previously, we have shown that the Smad2 and Smad3 genes function cooperatively during liver morphogenesis. Here we show that Smad2 and Smad3 are required at a full dosage for normal embryonic development. Animals lacking one allele of each gene exhibit a variably penetrant phenotype in which structures in the anterior and ventral midline are reduced or lost; additionally, we demonstrate that this craniofacial defect and the previously reported hepatic phenotypes are both due to defects in the definitive endoderm. A reduction of endodermal gene expression as well as a failure to displace the visceral endoderm occurs despite the formation of a normal foregut pocket. This precedes any defects in anterior patterning and likely causes the abnormalities observed in craniofacial and midline development, as well as hepatogenesis.

Animals↗

PubMed2004

Sox17 and beta-catenin cooperate to regulate the transcription of endodermal genes.

Recent studies have led to a model of the molecular pathway that specifies the endoderm during vertebrate gastrulation. The HMG box transcription factor Sox17 is a key component of this pathway and is essential for endoderm formation; however, the molecular events controlled by Sox17 are largely unknown. We have identified several direct transcriptional targets of Sox17, including Foxa1 and Foxa2. We show that beta-catenin, a component of Wnt signaling pathway, physically interacts with Sox17 and potentiates its transcriptional activation of target genes. We identify a motif in the C terminus of Sox17, which is conserved in all the SoxF subfamily of Sox proteins, and this motif is required for the ability of Sox17 to both transactivate target genes and bind beta-catenin. Nuclear beta-catenin is present in endoderm cells of the gastrula, and depletion of beta-catenin from embryos results in a repression of Sox17 target genes. These data suggest that in a mechanism analogous to Tcf/Lef interacting with beta-catenin, Sox17 and beta-catenin interact to transcribe endodermal target genes.

Amino Acid Motifs↗

PubMed2004

Disruption of genes encoding predicted inner arm dynein heavy chains causes motility phenotypes in Tetrahymena.

The multi-dynein hypothesis [Asai, 1995: Cell Motil Cytoskeleton 32:129-132] states: (1) there are many different dynein HC isoforms; (2) each isoform is encoded by a different gene; (3) different isoforms have different functions. Many studies provide evidence in support of the first two statements [Piperno et al., 1990: J Cell Biol 110:379-389; Kagami and Kamiya, 1992: J Cell Sci 103:653-664; Gibbons, 1995: Cell Motil Cytoskeleton 32:136-144; Porter et al., 1996: Genetics 144:569-585; Xu et al., 1999: J Eukaryot Microbiol 46:606-611] and there is evidence that outer arms and inner arms play different roles in flagellar beating [Brokaw and Kamiya, 1987: Cell Motil. Cytoskeleton 8:68-75]. However, there are few studies rigorously testing in vivo whether inner arm dyneins, especially the 1-headed inner arm dyneins, play unique roles. This study tested the third tenet of the multi-dynein hypothesis by introducing mutations into three inner arm dynein HC genes (DYH8, 9 and 12) that are thought to encode HCs associated with 1-headed inner arm dyneins. Southern blots, Northern blots, and RT-PCR analyses indicate that all three mutants (KO-8, 9, and 12) are complete knockouts. Each mutant swims slower than the wild-type cells. The beat frequency of KO-8 cells is lower than that of the wild-type cells while the beat frequencies of KO-9 and KO-12 are not different from that of wild-type cells. Our results suggest that each inner arm dynein HC is essential for normal cell motility and cannot be replaced functionally by other dynein HCs and that not all of the 1-headed inner arm dyneins play the same role in ciliary motility. Thus, the results of our study support the multi-dynein hypothesis [Asai, 1995: Cell Motil Cytoskeleton 32:129-132].

Animals↗

PubMed2003

Cardiac T-box factor Tbx20 directly interacts with Nkx2-5, GATA4, and GATA5 in regulation of gene expression in the developing heart.

Tbx20 is a member of the T-box transcription factor family expressed in the forming hearts of vertebrate and invertebrate embryos. We report here analysis of Tbx20 expression during murine cardiac development and assessment of DNA-binding and transcriptional properties of Tbx20 isoforms. Tbx20 was expressed in myocardium and endocardium, including high levels in endocardial cushions. cDNAs generated by alternative splicing encode at least four Tbx20 isoforms, and Tbx20a uniquely carried strong transactivation and transrepression domains in its C terminus. Isoforms with an intact T-box bound specifically to DNA sites resembling the consensus brachyury half site, although with less avidity compared with the related factor, Tbx5. Tbx20 physically interacted with cardiac transcription factors Nkx2-5, GATA4, and GATA5, collaborating to synergistically activate cardiac gene expression. Among cardiac GATA factors, there was preferential synergy with GATA5, implicated in endocardial differentiation. In Xenopus embryos, enforced expression of Tbx20a, but not Tbx20b, led to induction of mesodermal and endodermal lineage markers as well as cell migration, indicating that the long Tbx20a isoform uniquely bears functional domains that can alter gene expression and developmental behaviour in an in vivo context. We propose that Tbx20 plays an integrated role in the ancient myogenic program of the heart, and has been additionally coopted during evolution of vertebrates for endocardial cushion development.

Alternative Splicing↗

PubMed2003

Solubilization of acyl heterogeneous triacylglycerol in phosphatidylcholine vesicles.

The amount of acyl heterogeneous triacylglycerol (TG(HET)) solubilized by phosphatidylcholine (PC) vesicles, prepared by co-sonication of egg PC and small amounts (<6% w/w) of TG(HET), was determined using (13)C nuclear magnetic resonance (NMR). The acyl chains of TG(HET) were predominantly 16 or 18 carbons in length, 50% saturated, and approximately 21.7% (13)C isotopically enriched at the carbonyl carbon. The (13)C NMR spectra revealed two carbonyl resonances at chemical shift values between PC carbonyls and oil-phase TG carbonyls, confirming the presence of TG(HET) solubilized in PC vesicles. Oil-phase TG carbonyl peaks were present only in spectra of vesicles containing >3 wt % TG(HET). Integration of TG(HET) carbonyl resonances determined that PC vesicles solubilized 3.8 wt % of TG(HET), compared to 2.8 wt % of acyl homogeneous triolein. The difference between the maximum solubility of TG(HET) and that of homogeneous TG (TG(HOM)) with similar acyl chain lengths provides evidence that specific acyl composition, in addition to the acyl chain length of triacylglycerols, affects the solubility of TG in PC vesicles and TG-rich lipoprotein surfaces. Thus, TG(HET) may innately be a better model substrate than TG(HOM) for determination of substrate availability of TG at lipoprotein surfaces.

Liposomes↗