A molecular, phylogenetic and functional study of the <i>dADAR</i>mRNA truncated isoform during <i>Drosophila</i>embryonic development reveals an editing-independent function

Adenosine Deaminases Acting on RNA (ADARs) have been studied in many animal phyla, where they have been shown to deaminate specific adenosines into inosines in duplex mRNA regions. In Drosophila, two isoform classes are encoded, designated full-length (contains the editase domain) and truncated (lacks this domain). Much is known about the full-length isoform, which plays a major role in regulating functions of voltage-gated ion channel proteins in the adult brain. In contrast, almost nothing is known about the functional significance of the truncated isoform. In situ hybridization shows that both isoform mRNA classes are maternally derived and transcripts for both localize primarily to the developing central nervous system. Quantitative RT-PCR shows that about 35% of all dADAR mRNA transcripts belong to the truncated class in embryos. 3’-RACE results show that abundance of the truncated isoform class is developmentally regulated, with a longer transcript appearing after the mid-blastula transition.3’-UTR sequences for the truncated isoform have been determined from diverse Drosophila species and important regulatory regions including stop codons have been mapped. Western analysis shows that both mRNA isoform classes are translated into protein during embryonic development, as full-length variant levels gradually diminish. The truncated protein isoform is present in every Drosophila species studied, extending over a period spanning about 40 x 106 years, implying a conserved function. Previous work has shown that a dADAR protein isoform binds to the evolutionarily conserved rnp-4f pre-mRNA stem-loop located in the 5’-UTR to regulate splicing, while no RNA editing was observed, suggesting the hypothesis that it is the non-catalytic truncated isoform which regulates splicing. To test this hypothesis, we have utilized RNAi technology, the results of which support the hypothesis. These results demonstrate a novel, non-catalytic function for the truncated dADAR protein isoform in Drosophila embryonic development, which is very likely evolutionarily conserved.

Regulation of Expression for the RNP-4F Splicing Assembly Factor in the Fruit-Fly <i>Drosophila melanogaster</i>

Intron splicing in eukaryotic organisms requires the interactions of five snRNAs and numerous different proteins in the spliceosome. Although the molecular mechanism behind splicing has been well studied, relatively little is known about regulation of expression for these splicing factor proteins. One of these proteins is the evolutionarily-conserved Drosophila RNP-4F splicing assembly factor. This protein is transcribed from a single gene into two developmentally regulated mRNAs that differ in their 5’-UTR structure. In the longer isoform, known to be abundant in the developing fly central nervous system, a conserved retained intron which folds into a stem-loop has been implicated in expression control of the mRNA. Here, we describe construction and utilization of several new rnp-4f gene expression study vectors using a GFP reporter in the ΦC31 system. The results confirm our previous observation that presence of the regulatory stem-loop enhances RNP-4F protein expression. However, in that study, the enhancement factor protein was not identified. We show here that overexpression of the RNP-4F transgene compared to the control results in additional translation, as indicated by the GFP reporter in the fluorescent images. These results are interpreted to show that RNP-4F protein acts back on its own mRNA 5’-UTR regulatory region via a feedback pathway to enhance protein synthesis in the developing fly central nervous system. A model is proposed to explain the molecular mechanism behind rnp-4f gene expression control.

A phylogenetic study of <i>Drosophila</i>splicing assembly chaperone RNP-4F associated U4-/U6-snRNA secondary structure

The rnp-4f gene in Drosophila melanogaster encodes nuclear protein RNP-4F. This encoded protein is represented by homologs in other eukaryotic species, where it has been shown to function as an intron splicing assembly factor. Here, RNP-4F is believed to initially bind to a recognition sequence on U6-snRNA, serving as a chaperone to facilitate its association with U4-snRNA by intermolecular hydrogen bonding. RNA conformations are a key factor in spliceosome function, so that elucidation of changing secondary structures for interacting snRNAs is a subject of considerable interest and importance. Among the five snRNAs which participate in removal of spliceosomal introns, there is a growing consensus that U6-snRNA is the most structurally dynamic and may constitute the catalytic core. Previous studies by others have generated potential secondary structures for free U4-and U6-snRNAs, including the Y-shaped U4-/U6-snRNA model. These models were based on study of RNAs from relatively few species, and the popular Y-shaped model remains to be systematically re-examined with reference to the many new sequences generated by recent genomic sequencing projects. We have utilized a comparative phylogenetic approach on 60 diverse eukaryotic species, which resulted in a revised and improved U4-/U6-snRNA secondary structure. This general model is supported by observation of abundant compensatory base mutations in every stem, and incorporates more of the nucleotides into base-paired associations than in previous models, thus being more energetically stable. We have extensively sampled the eukaryotic phylogenetic tree to its deepest roots, but did not find genes potentially encoding either U4-or U6-snRNA in the Giardia and Trichomonas data-bases. Our results support the hypothesis that nuclear introns in these most deeply rooted eukaryotes may represent evolutionary intermediates, sharing characteristics of both group II and spliceosomal introns. An unexpected result of this study was discovery of a potential competitive binding site for Drosophila splicing assembly factor RNP-4Fto a5’-UTR regulatory region within its own pre-mRNA, which may play a role in negative feedback control.

空间飞行微重力降低裸藻光合电子传递并改变光合能量分配（英文）

利用神舟8号飞船的SIMBOX发射机会,对真实微重力影响裸藻光合作用活性进行了研究.我们发现,微重力降低了光合活性(Fv/Fm),提高了细胞内叶绿素a和胡萝卜素含量.快速叶绿素荧光动力学研究显示微重力降低了叶绿素荧光强度,但快速叶绿素荧光动力学曲线的形状(O-J-I-P)没有改变.在微重力处理下裸藻的最大光化学效率(φPo)、用于电子传递的量子产额(φEo)和光合作用性能指数(PIABS and PICS)都明显降低,但单位反应中心吸收的光能(ABS/RC)和单位反应中心耗散的能量(DIo/RC)都明显升高.77K低温荧光光谱实现微重力改变了能量在PSⅠ和PSⅡ之间的分配并出现了红移现象.这些结果表明真实微重力降低光合作用的活性有可能通过两个途径,即抑制裸藻抑制光合电子传递中PSⅡ的受体端和改变PSⅠ的结构从而引起流向PSⅠ的能量传递减少.

Risk Factors Associated with Preterm Birth at Hasan Sadikin General Hospital in 2015

Background: Preterm birth defined as childbirth occurring at less than 37 completed weeks or 259 days of gestation. The causes of preterm birth are complex and multifactorial, many risk factors that contribute in it. Knowledge of risk factors is crucial for predicting the incidence of preterm births. This study aimed to determine the factors associated with preterm birth at the Hasan Sadikin General Hospital. Method: This study was a cross-sectional analytic using secondary data. Data derived from medical records maternity patients in the Department of Obstetrics and Gynecology Hasan Sadikin General Hospital taken in 2015. 1944 patients’ medical records who gave birth met the inclusion criteria. Data analyses used were bivariate (chi square) and multivariate (logistic regression). Result: The result showed that the variables including age (p = 0.043, OR = 1.586), antenatal care (p p p p p p Conclusion: There is a significant relationship between age, antenatal care, preterm birth history birth, anemia, hypertension, and antepartum hemorrhage with preterm birth as risk factors.

The Heme-Binding Protein SOUL3 of Chlamydomonas reinhardtii Influences Size and Position of the Eyespot

The flagellated green alga Chlamydomonas reinhardtii has a primitive visual system, the eyespot. It is situated at the cells equator and allows the cell to phototax. In a previous proteomic analysis of the eyespot, the SOUL3 protein was identified among 202 proteins. Here, we investigate the properties and functions of SOUL3. Heterologously expressed SOUL3 is able to bind specifically to hemin. In C. reinhardtii, SOUL3 is expressed at a constant level over the diurnal cycle, but forms protein complexes that differ in size during day and night phases. SOUL3 is primarily localized in the eyespot and it is situated in the pigment globule layer thereof. This is in contrast to the channelrhodopsin photoreceptors, which are localized in the plasma membrane region of the eyespot. Knockdown lines with a significantly reduced SOUL3 level are characterized by mislocalized eyespots, a decreased eyespot size, and alterations in phototactic behavior. Mislocalizations were either anterior or posterior and did not affect association with acetylated microtubules of the daughter four-membered rootlet. Our data suggest that SOUL3 is involved in the organization and placement of the eyespot within the cell.