RESUMO
Rho family GTPases are thought to regulate actin-dependent processes, but their functions in vivo are still poorly understood. We have investigated the function of a new, widely expressed Rho family member in C. elegans by analyzing mutations in the endogenous gene. Activated and null alleles all inhibit cell migration, demonstrating that this protein is required for cell migration in vivo. Only a small subset of the migrations inhibited by activating mutations are inhibited by null mutations, suggesting that considerable functional redundancy exists within this system. Our findings support this conclusion and show that mig-2 functions redundantly with another pathway to regulate nuclear migration. Surprisingly, activated alleles also cause misguided axon growth, suggesting that Rho family GTPases may couple guidance cues to process outgrowth.
Assuntos
Axônios/fisiologia , Caenorhabditis elegans/genética , Movimento Celular/fisiologia , GTP Fosfo-Hidrolases/genética , Proteínas de Ligação ao GTP/genética , Neurônios/citologia , Alelos , Animais , Animais Geneticamente Modificados , Axônios/química , Caenorhabditis elegans/química , Caenorhabditis elegans/citologia , Membrana Celular/química , Clonagem Molecular , GTP Fosfo-Hidrolases/análise , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Helminto/genética , Dados de Sequência Molecular , Mutação/fisiologia , Neurônios/química , Neurônios/ultraestrutura , Homologia de Sequência de AminoácidosRESUMO
Previously, a small region of the 3'-untranslated region (3' UTR) of Xlhbox2B mRNA was shown to be sufficient for sequence-specific endonucleolytic cleavage after injection into Xenopus oocytes. Here, we report an in vitro RNA degradation reaction that mimics the in vivo result accurately. The reaction also reveals that oocytes contain a sequence-specific RNA-binding factor that inhibits the endoribonuclease. These opposing activities may be regulated during Xenopus oogenesis. Partial purification shows that the endonuclease does not require translation or ribosomes and does not resemble previously described RNA processing complexes. We have isolated another Xenopus cDNA, Xoo1, that contains a long, repetitive destabilizing element similar to the one in Xlhbox2B. Based on a comparison of these natural destabilizing sequences and in vitro mutagenesis experiments, we find that a single destabilizing site is, at most, 19 bases in length and that the endonuclease and protective factor recognition sites may be overlapping subsets of this sequence. Finally, we show that Drosophila embryos contain similar activities, each of which can use Xenopus recognition sites. This level of conservation suggests an important biological function for this system of post-transcriptional regulation.
Assuntos
Endorribonucleases/metabolismo , Processamento Pós-Transcricional do RNA , RNA Mensageiro/metabolismo , Animais , Sequência de Bases , Sítios de Ligação , Ligação Competitiva , Sequência Conservada , Drosophila melanogaster/genética , Embrião não Mamífero/enzimologia , Meia-Vida , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Oócitos/enzimologia , RNA Mensageiro/química , Sequências Repetitivas de Ácido Nucleico , Mapeamento por Restrição , Análise de Sequência de RNA , Especificidade por Substrato , Xenopus laevis/genéticaRESUMO
A study was conducted to determine the values of several chemical components of dental cementum. Extracted teeth from continuous residents of low, optimal, and high natural fluoride areas were selected to determine the effects of fluoride exposure and age on the fluoride, citrate and carbonate content of cervical and apical areas of sound and periodontally diseased human cementum. 1. For normal cementum the mean fluoride content of all 18 groups was 0.443 plus or minus 0.04%on an ash basis and 0.253 plus or minus 0.02% on a dry weight basis; the mean citrate content was 1.52 plus or minus 0.10% on an ash basis and 0.86 plus or minus 0.06% on a dry weight basis; the mean carbonate content was 4.95 plus or minus 0.39% on an ash basis and 2.79 plus or minus 0.22% on a dry weight basis. 2. For diseased cementum the mean fluoride content of all 12 groups was 0.575 plus or minus 0.03% on an ash basis and 0.342 plus or minus 0.02% on a dry weight basis; the mean citrate content was 1.19 plus or minus 0.07% on an ash basis and 0.71 plus or minus 0.05% on a dry weight basis; the mean carbonate content was 5.95 plus or minus 0.47% on an ash basis and 3.50 plus or minus 0.28% on a dry weight basis. 3. There is a general increment in fluoride content of normal cervical and apical cementum, both with age and fluoride exposure. For diseased cementum this held true for fluoride exposure but not age. 4. The mean fluoride content of cervical cementum is greater than that of apical cementum for both normal and diseased groups. For citrate and carbonate no definite statistical differences were noted between cervical and apical areas. 5. The diseased cementum has a significantly higher mean fluoride content, a significantly lower citrate content, and a slightly, but not statistically significant, higher carbonate content than normal cementum. 6. Unlike bone, no inverse fluoride/citrate or fluoride/carbonate relationship was observed. 7. For both normal and diseased groups the carbonate content of cementum of individuals over age 40 tended to be lower than for individuals 20-40 years of age.
