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Developmental regulation of cmp1, a gene encoding a multidomain conidiospore surface protein of Trichoderma.

A gene encoding a developmentally regulated polypeptide of Trichoderma (strain ATCC 32173) was isolated, with the help of an antibody against a 62-kDa protein whose abundance strongly increases during photoinduced sporulation. The amino acid sequence deduced from this gene, cmp1 (conidial multidomain protein), is a 135-kDa polypeptide consisting of several domains. Although reminiscent of known structural modules, two of the domains may define novel families. The protein is apparently processed to give the 62-kDa species. Immunogold labeling electron microscopy localized the antigen to the membrane or inner wall layers. The mRNA is strongly up-regulated during sporulation. At least part of this regulation is likely to be conferred by several elements identified in the upstream region, with homology to elements recognized by fungal transcription factors for regulation by conidiation, light, and nitrogen stress. The developmental regulation, cell surface location, and modular structure suggest a function in cell-cell interactions, detection of the wall by the cell, or anchoring of the plasma membrane to the wall.

Amino Acid Sequence↗

Expression of a novel member of the ATP1G1/PLM/MAT8 family, phospholemman-like protein (PLP) gene, in the developmental process of mouse cerebellum.

We have identified a new member of the ATP1G1/PLM/MAT8 family, named phospholemman-like protein (PLP), from a mouse cerebellum cDNA library. The family consists of small transmembrane proteins that modulate the activities of some ion channels. The deduced amino acid sequence of PLP consists of 93 residues that contain the ATP1G1/PLM/MAT8 motif and a single transmembrane domain, and is most similar to the sequence of mouse phospholemman. In situ hybridization analysis showed that the PLP gene is highly expressed in cerebellar granule cells. PLP expression is elevated in the postnatal developing cerebellum. Thus, it may be implicated in the proliferation, differentiation, and axon elongation of granule cells as they mature and migrate to the internal granule layer.

Amino Acid Sequence↗

Developmental regulation of rat brain/Hep G2 glucose transporter gene expression.

The developmental regulation of rat brain-derived/Hep G2 glucose transporter gene expression was studied by means of Northern blot hybridization, using a rat brain glucose transporter cDNA probe, in order to directly quantify steady state glucose transporter mRNA levels. The results obtained showed different tissue-specific patterns of glucose transporter mRNA levels during ontogenesis; while in brain there was a sustained increase in the levels of the message from 20 days embryogenesis until 50 days postnatal, other organs such as heart, lung, liver, and muscle expressed maximal levels of the glucose transporter mRNA in 20-day fetuses and 1-day neonates, decreasing subsequently to very low levels. The relative expression of the glucose transporter mRNA in the different tissues, at both fetal and adult stages, was analyzed using a solution hybridization-RNase protection assay. This approach revealed that, while the heart expresses the highest levels of glucose transporter mRNA at 20 days of fetal life, the brain shows the highest levels at the adult stage. These results indicate a tissue-specific ontogenic pattern of glucose transporter gene expression, suggesting a developmental role for this glucose transporter gene product.

Animals↗

Expression of novel lipocalin-like milk protein gene is developmentally-regulated during lactation in the tammar wallaby, Macropus eugenii.

We have identified a novel whey protein (late lactation protein B; LLPB) that is first secreted in the milk of the tammar wallaby around day 200 of lactation. The LLPB cDNA clone of 843 base pairs encodes a mature protein of 156 amino acids. LLPB shares 65 and 48% nucleotide and deduced amino acid identity, respectively, with a previously identified late lactation protein A (LLPA). Both these proteins share significant amino acid sequence homology with the lipocalin protein family. Expression of the LLPB gene is induced between days 200 and 240 of lactation, in contrast to expression of the LLPA gene, which is induced at around 145 days of lactation. Maximal expression of both genes in mammary explants from tammars at 213 days of lactation required a combination of prolactin, insulin and hydrocortisone. Transcripts of LLPA, LLPB and beta-lactoglobulin (TBLG) were localized to the same cells by in situ hybridization. A substantial level of alveolar maturation is required for expression of the LLP genes, unlike TBLG, which is expressed in immature alveoli. We hypothesize that the temporal expression of the LLPB and LLPA genes may be regulated both by endocrine stimuli and factors intrinsic to the mammary gland.

Amino Acid Sequence↗

Association of early sporulation genes with suggested developmental decision points in Streptomyces coelicolor A3(2).

Cytological analysis of a series of Streptomyces coelicolor A3(2) mutants with disruptions of early sporulation (whi, for white aerial mycelium) genes in an isogenic background has provided new information about the role of whiH, and confirmed and extended previous knowledge about whiG, whiA and whiB. The characteristic straight aerial hyphae of whiG mutants contained normally spaced vegetative-like septa, while mutants in whiA or whiB had abnormally long and coiled aerial hyphae almost devoid of septation. whiG, whiA and whiB were all absolutely required for sporulation septation, and for all visible signs of nucleoid condensation and partitioning and other changes associated with later stages of sporulation. On the other hand, whiH appeared to enhance low basal levels of these processes. Thus, whiH mutant aerial hyphae were divided into loosely coiled fragments of variable sizes by what appeared to be a few sporulation septa. These fragments showed some spore-like characteristics and contained condensed and aberrantly partitioned nucleoids. whiG, whiA and whiB were epistatic to whiH on the criterion that they prevented such fragments from forming in double mutants. These spore-like features and the synthesis of clearly detectable levels of the whiE-directed grey spore pigment were not due to any residual activity of previously studied whiH alleles since they were retained by a constructed whiH null mutant. A model is presented that explains the mutant phenotypes by proposing two early developmental decision points involved in commitment to sporulation septation, one requiring whiG and the other requiring whiA and whiB.

Alleles↗

Evolution of the response patterns to dietary carbohydrates and the developmental differentiation of gene expression of alpha-amylase in Drosophila.

Intraspecific variation of alpha-amylase activity in D. melanogaster and D. immigrans, which is distantly related to D. melanogaster, and interspecific variation of alpha-amylase activity in 18 Drosophila species were examined. The amount of intraspecific variation of alpha-amylase activities measured in terms of coefficient of variation in D. melanogaster and D. immigrans was one-half and one-tenth or less, respectively, of the interspecific variation in 18 Drosophila species. We also surveyed the response patterns of alpha-amylase activity to dietary carbohydrates at the larval and adult stages. The levels of alpha-amylase activity depended on both repression by dietary glucose (glucose repression) and induction by dietary starch (starch induction). In general, our data suggest that glucose repression was conserved among species at both stages while starch induction was mainly observed in larvae, although the degree of the response depended on species. In D. lebanonensis lebanonensis and D. serrata, larvae expressed electrophoretically different alpha-amylase variants (isozymes) from those of adult flies. These results may suggest that the regulatory systems responsible both for the response to environment and developmental expression are different among species in Drosophila.

Animals↗

Developmental behavior of gene expression for brown rice thickness under different environments.

The dynamic changes of genetic effects, including main effects, and genotype x environment (GE) interaction effects on brown rice thickness (BRT) across environments were investigated by using the developmental genetic models. Seven cytoplasmic male sterile lines of indica rice (Oryza sativa L.) as females and five restoring lines as males were used in a factorial design to produce grains of F(1)s and F(2)s in two environments (years) for developmental genetic analysis. The results indicate that genetic effects, especially GE interaction effects of triploid endosperm genes, cytoplasm genes, and diploid maternal plant genes were important to the performance of BRT at various filling stages of rice. The BRT was genetically controlled by the net genetic effects of genes expressed at the early and late filling stages (1-7 days and 15-21 days after flowering, respectively). The differences in net genetic effects under different environments for endosperm, cytoplasm, and maternal plant genes were found, and the net GE interaction effects were more important to BRT at the early filling and mature stages of rice. Some net genetic effects, especially for net cytoplasm effects spasmodically expressed, were detected among filling stages. Higher additive and cytoplasm main effects, along with their interaction effects, were found, which would be useful for selection for BRT in breeding programs. The predicated genetic effects at different filling stages show that the parents of V20 and Xieqingzao were better than others for improving BRT of progenies.

Analysis of Variance↗

Murine pulmonary myocardium: developmental analysis of cardiac gene expression.

Long-standing observations that cardiac muscle exists in the walls of the pulmonary and caval veins have recently been confirmed at the molecular level (Lyons et al. [1990] J. Cell Biol. 111:2427-2436; Springall et al. [1988] Thorax 43:44-52; Subramaniam et al. [1991] J. Biol. Chem. 266:24613-24620). Using ventricle- and atrial-specific riboprobes, we determined that the pulmonary myocardium exhibits an atrial pattern of cardiac-specific gene expression. Additionally, the developmental pattern of expression was studied using a riboprobe specific to the alpha-cardiac myosin heavy chain (alpha-MHC) gene transcript. We find that alpha-MHC gene expression is first detectable in the lung between 13.9-14.3 days post-coitum. Extension of the alpha-MHC specific hybridization signal into the pulmonary venous bed progresses through the neonatal period. The data are consistent with the hypothesis that the extension of alpha-MHC gene expression into the lung occurs via the migration of atrial myoblasts into the vein during atrial septation and remodeling of the sinus venosus and pulmonary venous trunk.

Animals↗

Structure, organization and evolution of developmentally regulated chorion genes in a silkmoth.

We describe in detail two cloned chromosomal DNA segments from Antheraea polyphemus, each bearing multiple chorion genes. Two types of genes are found per segment, each in two or three copies: they belong to both major chorion multigene families (A and B), but are expressed during the same developmental period (middle choriongenesis in one segment, late in the other). A and B genes alternate and are paired, lie in divergent transcriptional orientation and are in very close proximity to each other (mRNA cap sites are separated by a 5' flanking sequence 264 or 325 bp long). Each AB pair is embedded within a large, tandemly repeating unit. Within that unit, the homologous 3' flanking sequences that separate gene pairs evolve rapidly and are frequently interrupted by long segments representing inserts (or deletions). These segments would decrease unequal crossing-over, facilitating rapid evolution of the chorion multigene families. A total of 10.4 kb of DNA has been sequenced, permitting detailed comparisons of genes, their introns, 5' flanking and immediately 3' flanking regions. Genes range over two orders of sequence similarity, from highly releated to very disparate (gene copies; members of the same multigene family; members of different families). Among their universal features are a common cap-site sequence, a single intron invariably located at the same position within the signal peptide-encoding region, and a Hogness box 21 to 23 nucleotides upstream from the cap site. Features of possible paired promoters occupying the short 5' flanking region are discussed. Genes evolve both by base substitutions and by segmental mutations, which are almost invariably deletions/insertions related to small, direct repeats.

Animals↗

Developmentally regulated Drosophila gene family encoding the fork head domain.

We have isolated seven Drosophila genes by means of low-stringency hybridization to a DNA probe containing the coding sequence for the protein domain shared by the rodent hepatocyte-enriched nuclear transcription factor HNF3A (alpha) and the product of the Drosophila region-specific homeotic gene fork head (fkh). The previously unreported genes encode a 110-amino acid conserved sequence, which we call the fork head (fkh) domain. Two of these fkh-domain-encoding genes ("FD genes") map to the sloppy paired locus (slp), which exerts segmentation gene function. The expression patterns of the other FD genes suggest that their protein products are likely to be involved in gut formation, mesoderm specification, and some specific aspects of neural development. The FD gene products presumably represent a family of transcription factors that, like the previously identified DNA-binding proteins, contribute to early developmental decisions in cell fates during embryogenesis.

Amino Acid Sequence↗

Disruption of glycogen phosphorylase gene expression in Dictyostelium: evidence for altered glycogen metabolism and developmental coregulation of the gene products.

Glycogen phosphorylase 1 and 2, the isozymes responsible for glycogen degradation, are encoded by separate genes in Dictyostelium. The two gene products display different transcriptional and translational expression and distinct post-translational regulation. Using DNA-mediated transformation, Dictyostelium clones which lacked either glycogen phosphorylase 1 or 2 (gp1 or gp2) expression were obtained. The loss of either enzyme did not change axenic growth patterns, developmental progression, or gross organismic morphology. In gp1- strains, glycogen accumulated to a 17- to 28-fold higher level during late stationary phase without any obvious detrimental effects. This implies that no alternative pathway for glycogen degradation is present in amoebae, and that glycogen metabolism is not critical for vegetative cell growth. Developmental glycogen concentrations were not altered significantly in any of the transformants, but in gp2- cells the posttranslational regulation of the intact gp1 enzyme was apparently modulated to compensate for the loss of gp2. Western blots of microdissected, lyophilized Dictyostelium slugs and early culminates showed that gp2 was found in both prestalk and prespore cells, with a slight enrichment in prespore cells. The gp1 protein was highly enriched in prestalk cells in the parental strain. In gp2- transformants, however, gp1 was detected in equal amounts in both cell types. The loss of gp2 led to a shift in the cell-type-specific expression pattern of gp1, presumably due to developmental coordinate regulation of gp1 and gp2 at the translational and/or transcriptional level.

Animals↗

Nonuniform evolution of duplicated, developmentally controlled chorion genes in a silkmoth.

We report the sequence of A/B.L1, a tightly linked pair of genes from the A and B chorion families in Bombyx mori. Comparison with the previously characterized A/B.L11 and A/B.L12 pairs from the same species reveals moderate sequence divergence, which is clearly nonuniform. Although the average divergence of A/B.L12 from the other two pairs is more than double that between A/B.L11 and A/B.L1, the ratio differs by more than 30-fold in different DNA regions. One domain of the A gene is highly divergent between A/B.L12 and A/B.L1 or A/B.L11, but essentially invariable in the latter two. In well-aligned DNA segments, nearly all mutated sites (111/112) show variants shared by two of the three sequences, in 42% of the cases between the more distantly related pairs (A/B.L12 and either A/B.L1 or A/B.L11). Eight of the variants shared by distantly related pairs are clustered within 51 bp, suggesting the possibility that they arose through sequence transfers between gene pairs, rather than being primitive or resulting from independent mutations. The short intergenic, putatively regulatory DNA of the developmentally middle A/B.L1 and A/B.L11 pairs resembles that of the late HcA/HcB pairs, particularly in patches that may correspond to cis-regulatory elements.

Animals↗

cDNA cloning and developmental alterations in gene expression of the two Pit-1/GHF-1 transcription factors in the chicken pituitary.

Pit-1/GHF-1 (Pit-1) transcription factors promote the gene expressions for growth hormone (GH), prolactin (PRL), and the beta chain of thyroid-stimulating hormone in vertebrate pituitary glands. The present study analyzed the nature of chicken Pit-1s (cPit-1s) and their developmental expressions in the pituitary. Chicken pituitary expressed two cPit-1 mRNAs encoding cPit-1alpha and cPit-1gamma composed of 335 and 327 amino acid residues, respectively. They possessed different N-terminal regions and the common C-terminal regions containing a POU-specific domain and a POU homeodomain. Northern blot analysis revealed the pituitary-specific expressions of these Pit-1 mRNAs, and the Pit-1alpha mRNA expressions were two to three times higher than those for Pit-1gamma in both cephalic and caudal lobes of the pituitary. The cPit-1alpha and gamma mRNA expressions simultaneously increased after hatching until 4 weeks and then slightly decreased at 5 weeks. Similar gene expression profiles were observed for GH and PRL during the posthatch developmental period.

Amino Acid Sequence↗

Selective developmental regulation of gene expression for insulin-like growth factor-binding proteins in mouse spinal cord.

STUDY DESIGN: Prospective, randomized experimental study in mice. STUDY OBJECTIVE: To determine whether insulin-like growth factor binding proteins (IGFBPs) are present in mouse spinal cord and, if so, what role they play in its development. SUMMARY OF BACKGROUND DATA: Insulin-like growth factors are well recognized hormonal effectors of growth hormone and are expressed in the mammalian spinal cord. The IGFBPs are a group of six genetically distinct proteins that bind IGFs and modulate their bioactivity. They appear in the brain during development, localize to the neuromuscular junction, and promote motor neuron survival. The benefit of IGF-I in amyotrophic lateral sclerosis ALS and its potential use in preventing motor neuron apoptosis in spinal cord injury dictates that studies of the presence and response of IGFBPs in that tissue be performed. METHODS: The IGFBPs in mouse spinal cord were analyzed by Western ligand blot, Western immunoblot, and reverse transcription-polymerase chain reaction at various time points from embryonic day 14 to postnatal day 30. RESULTS: Three IGFBPs with molecular masses of 24, 28, and 32 kDa were found, the latter two being the most prominent. The data indicate that these are IGFBP-4, -5, and -2. CONCLUSION: Both IGFBP-2 and BP-5 are developmentally regulated in mouse spinal cord, with higher levels of those at early embryonic stages indicating their potential role in development of the mouse spinal cord.

Animals↗

Aedes aegypti peroxidase gene characterization and developmental expression.

The functions of insect peroxidases include detoxification, stabilization of extracellular matrices, and possible involvement in insect immunity. The current study describes the isolation of a peroxidase gene, AePox, and its cDNA from the mosquito, Aedes aegypti. AePox codes for a protein that is homologous to various heme-peroxidases from vertebrates and invertebrates, with highest identity to Drosophila melanogaster peroxidase (62%). Sequence comparison identified several functionally and structurally conserved domains in the mosquito peroxidase, including a heme environment, a calcium binding site, and five possible disulfide bridges. These results imply that AePOX may likely have a similar structure and catalytic mechanism as those described for the mammalian myeloperoxidase superfamily. Expression studies demonstrate that AePox is transcribed in mosquito larvae and pupae, but not in adults, in ovaries, or in early embryos. However, AePOX protein is present in all mosquito stages and possibly has a maturation process that is similar to that of human myeloperoxidase. Unlike most human peroxidases, the AePox gene contains a TATA box and an ecdysone response element (EcRE).

Aedes↗

Comparison of transcription of multiple genes at three developmental stages of the plant pathogen Sclerotinia sclerotiorum.

The ascomycete Sclerotinia sclerotiorum is a plant pathogen with a very broad host range. In order to identify and characterize genes involved in S. sclerotiorum infection of Brassica napus (canola), expressed sequence tags (ESTs) were examined from libraries prepared from three tissues: complex appressorium (infection cushions), mycelia grown on agar and lesions formed on leaves of B. napus. A high proportion of genes (68%) had not been previously reported for S. sclerotiorum in public gene or EST databases. The types of novel genes identified in the infection cushion library highlights the functional specificity of these structures and similarities to appressoria in other fungal pathogens. Quantitative real-time PCR was used to analyse tissue specificity and timing of transcription of genes with best matches to MAS3 (appressoria-associated protein from Magnaporthe grisea), cellobiohydrolase I, oxaloacetate acetylhydrolase, metallothionein, pisatin demethylase, and an unknown gene with orthologs in fungal pathogens but not in saprophytic fungi.

Ascomycota↗

Eye evolution: a question of genetic promiscuity.

Animal eyes have long served as a classical example of independent origin followed by convergence of structures onto a few different solutions. During the past decade this view has been challenged by the discovery of shared developmental regulatory genes. The Pax6 gene in particular is almost universally employed for eye formation in bilaterian animals, despite widely different embryological origins. The resulting controversy on the multiple or single origins of animal eyes has gradually been sharpened by continuing discoveries of further general similarities in the genetic regulatory circuits of eye development. Recent work on gene expression in specified cell types, together with comparative studies of developmental genes in cnidarians, now show some promise to a solution of the controversy.

Animals↗

Cloning of chick cellular retinol-binding protein, type II and comparison to that of some mammals: expression of the gene at different developmental stages, and possible involvement of RXRs and PPAR.

We cloned chick cellular retinol-binding protein, type two (CRBP II) cDNA and compared it with those of some mammals. The deduced amino acid sequence showed that chick CRBP II was one amino acid greater in size than those of mammals, and the nucleotide sequence of chick CRBP II shared 72%-75% similarity with those of mammals. RNA blot hybridization analysis showed that CRBP II transcript of 0.7 kb was first detected in the duodenum of day-18 embryonic chick, and exhibited a rapid increase during 24 hr around the hatching. Northern blot hybridization also revealed that the transcripts of two types of retinoid X receptors (RXR alpha and RXR gamma) and peroxisome proliferator-activated receptor (PPAR) were expressed in the chick duodenum at hatching. The organ culture of day 16 embryonic chick duodenum showed that the addition of 9-cis retinoic acid in the medium caused a significant increase in CRBP II mRNA levels. In addition, arachidonic acid, from which putative ligands for PPAR were supposed to be generated, was accumulated around hatching in the duodenum. The results may suggest that the abrupt increase of the CRBP II gene expression in the chick duodenum around hatching may be related with RXRs and/or PPAR.

Animals↗