Four Ubiquitously Expressed Genes,RD(D6S45)–SKI2W(SKIV2L)–DOM3Z–RP1(D6S60E), Are Present between Complement Component Genes FactorBandC4in the Class III Region of the HLA

The association of the HLA class III region with many diseases motivates the investigation of unidentified genes in the 30-kb segment between complement component genesBfandC4. RD,which codes for a putative RNA binding protein, is 205 bp downstream ofBf. SKI2W(HGMW-approved symbol SKIV2L), a DEVH-box gene probably involved in RNA turnover, is 171 bp downstream ofRD(HGMW-approved symbol D6S45).RP1(HGMW-approved symbol D6S60E) is located 611 bp upstream ofC4.The DNA sequence between humanRDandRP1was determined and the exon–intron structure ofSKI2Welucidated.SKI2Wconsists of 28 exons. The putative RNA helicase domain of Ski2w is encoded by 9 exons. Further analysis of the 2.5-kb intergenic sequence betweenSKI2WandRP1led to the discovery ofDOM3Z.The full-length cDNA sequence ofDOM3Zencodes 396 amino acids with a leucine zipper motif. Dom3z-related proteins are present in simple and complex eukaryotes. InCaenorhabditis elegans,Dom3z-related protein could be involved in the development of germ cells. HumanRD–SKI2WandDOM3Z–RP1are arranged as two head-to-head oriented gene pairs with unmethylated CpG sequences at the common 5′ regulatory region of each gene pair. The ubiquitous expression pattern suggests that these four genes are probably housekeeping genes.     

Chromosomal Mapping ofTmp(Emp1),Xmp(Emp2), andYmp(Emp3), Genes Encoding Membrane Proteins Related toPmp22

We have recently characterized a novel mammalian gene family, encoding membrane glycoproteins with four trans-membrane domains. This gene family includes the previously studiedPMP22,which is involved in the Charcot–Marie–Tooth neuropathy, and three novel genes:TMP, XMP,andYMP(HGMW-approved symbolsEMP1, EMP2andEMP3,respectively). TheTmp(tumor-associated membrane protein) gene was isolated from a c-mycinduced mouse brain tumor and is expressed in several highly proliferative cell types. We have now isolated cDNAs of the mouseXmpandYmpgenes and determined the chromosomal localization of mouseTmp, Xmp,andYmp. Tmpwas mapped to mouse chromosome 6,Xmpwas mapped to chromosome 16, andYmpwas mapped to chromosome 7.TmpandYmpmap to paralogous chromosomal regions, whereasXmpmaps to a chromosomal region that is putatively paralogous to a region on chromosome 11, to whichPmp22was previously mapped. These data suggest that this family of membrane glycoproteins evolved as a result of chromosomal duplications.     

Chemokine PARC Gene (SCYA18) Generated by Fusion of Two MIP-1α/LD78α-like Genes

Two loci in the human genome, chromosomes 4q12–q21 and 17q11.2, contain clusters of CXC and CC chemokine subfamily genes, respectively. Since mice appear to contain fewer chemokine genes than humans, numerous gene duplications might have occurred in each locus of the human genome. Here we describe the genomic organization of the human pulmonary and activation-regulated CC chemokine (PARC), also known as DC-CK1 and AMAC-1. Despite high sequence similarity to a CC chemokine macrophage inflammatory protein-1α (MIP-1α)/LD78α, PARC is chemotactic for lymphocytes and not for monocytes and does not share its receptor with MIP-1α. Analyses of the BAC clones containing the humanPARCgene indicated that the gene is located most closely toMIP-1α(HGMW-approved symbolSCYA3) andMIP-1β(HGMW-approved symbolSCYA4) on chromosome 17q11.2. Dot-plot comparison suggested that thePARCgene had been generated by fusion of twoMIP-1α-like genes with deletion and selective usage of exons. Base changes accumulated before and after the fusion might have adapted the gene to a new function. Since there are variably duplicated copies of theMIP-1αgene calledLD78β(HGMW-approved symbolSCYA3L) in the vicinity of theMIP-1αgene, the locus surrounding theMIP-1αgene seems to be a “hot spring” that continuously produces new family genes. This evidence provides a new model, duplication and fusion, of the molecular basis for diversity within a gene family.     

CBFA2T1,a Gene Rearranged in Human Leukemia, Is a Member of a Multigene Family

MTG8(HGMW-approved symbolCBFA2T1) was originally identified as one of the loci involved in the t(8;21)(q22;q22) of acute myeloid leukemia. We characterize two humanMTG8-related genes,MTGR1andMTGR2(HGMW-approved symbolsCBFA2T2andCBFA2T3). The former is duplicated in mouse, one locus possibly being a retroposon. MultipleMTG8-related sequences are found in several vertebrate species, from fish to mammals, albeit not in a urodele.MTGR2maps to 16q24 and, likeMTG8andMTGR1,is close to one of three loci encoding a syntrophin (dystrophin-associated proteins). Moreover, an alternativeMTGR1promoter/5′ exon is contained within the α1-syntrophin locus. Thus, the two classes of genes may define novel paralogous groups.MTGR1is expressed mainly in brain, whileMTGR2is expressed in the thymus and possibly in monocytes. LikeMTG8, MTGR1is transcribed into a number of isoforms due to alternative splicing of different 5′ exons onto a common splice acceptor site. Comparison of the three predicted human MTG8-related polypeptides to theirDrosophilacounterpart (nervy) highlights four separate regions of sequence conservation that may correspond to distinct domains. The most NH₂-terminal of these is proportionately more conserved among the human polypeptides, presumably due to specific structural/functional constraints.     

Comparative Prey Suitability ofOstrinia nubilalisEggs andAcyrthosiphon pisumforColeomegilla maculata

The effects ofOstrinia nubilalis(Hübner) (Lepidoptera: Pyralidae) eggs andAcyrthosiphon pisum(Harris) (Homoptera: Aphididae), when provided as single prey species and in combination, on life history characteristics ofColeomegilla maculataDeGeer (Coleoptera: Coccinellidae) larvae and adults were quantified. Preimaginal development was not influenced by the larval prey regime; development at 26 ± 1°C was completed in approximately 13.5 days onO. nubilaliseggs,A. pisum,orA. pisumalternated daily withO. nubilaliseggs. The resulting adults weighed 13.0, 10.7, and 12.5 mg when reared onO. nubilaliseggs,A. pisum,andA. pisumalternated daily withO. nubilaliseggs, respectively. Eighteen percent of the individuals died when reared onA. pisum,28% died when reared onO. nubilaliseggs, and 22% died when fedA. pisumalternated daily withO. nubilaliseggs. Seven adult diet combinations, based on diet regimes of larvae and adults, did not cause significant differences in preoviposition period, interoviposition period, and the number of days on which eggs were laid. Total fecundity was influenced both by larval and adult diet. The diet that resulted in highly fecund females wasA. pisumalternated daily withO. nubilaliseggs for larvae andO. nubilaliseggs for adults. FemaleC. maculatafedO. nubilaliseggs had the highest intrinsic rate of increase and net reproductive rate.     

Molecular Cloning, Expression Pattern, and Chromosomal Localization of the Human Na–Cl Thiazide-Sensitive Cotransporter (SLC12A3)

Electrolyte homeostasis is maintained by several ion transport systems. Na–(K)–Cl cotransporters promote the electrically silent movement of chloride across the membrane in absorptive and secretory epithelia. Two kidney-specific Na–(K)–Cl cotransporter isoforms are known, so far, according to their sensitivity to specific inhibitors. We have cloned the human cDNA coding for the renal Na–Cl cotransporter selectively inhibited by the thiazide class of diuretic agents. The predicted protein sequence of 1021 amino acids (112 kDa) shows a structure common to the other members of the Na–(K)–Cl cotransporter family: a central region harboring 12 transmembrane domains and the 2 intracellular hydrophilic amino and carboxyl termini. The ex- pression pattern of the human Na–Cl thiazide-sensitive cotransporter (hTSC, HGMW-approved symbol SLC12A3) confirms the kidney specificity. hTSC has been mapped to human chromosome 16q13 by fluorescencein situhybridization. The cloning and characterization of hTSC now render it possible to study the involvement of this cotransport system in the pathogenesis of tubulopathies such as Gitelman syndrome.     

Genetic characterization of hydroxyurea-resistant mutants obtained from cell cultures of Nicotiana tabacum

Summary Genetic characterization of seven hydroxyurearesistant (HuR) mutants selected from cell cultures of Nicotiana tabacum var. Xanthi has shown that in each case resistance is inherited as a single, dominant, nuclear mutation. Four HuR mutants for which linkage analysis has been performed identify two unlinked loci.     

Chemokine PARC gene (SCYA18) generated by fusion of two MIP-1alpha/LD78alpha-like genes

Two loci in the human genome, chromosomes 4q12-q21 and 17q11.2, contain clusters of CXC and CC chemokine subfamily genes, respectively. Since mice appear to contain fewer chemokine genes than humans, numerous gene duplications might have occurred in each locus of the human genome. Here we describe the genomic organization of the human pulmonary and activation-regulated CC chemokine (PARC), also known as DC-CK1 and AMAC-1. Despite high sequence similarity to a CC chemokine macrophage inflammatory protein-1alpha (MIP-1alpha)/LD78alpha, PARC is chemotactic for lymphocytes and not for monocytes and does not share its receptor with MIP-1alpha. Analyses of the BAC clones containing the human PARC gene indicated that the gene is located most closely to MIP-1alpha (HGMW-approved symbol SCYA3) and MIP-1beta (HGMW-approved symbol SCYA4) on chromosome 17q11.2. Dot-plot comparison suggested that the PARC gene had been generated by fusion of two MIP-1alpha-like genes with deletion and selective usage of exons. Base changes accumulated before and after the fusion might have adapted the gene to a new function. Since there are variably duplicated copies of the MIP-1alpha gene called LD78beta (HGMW-approved symbol SCYA3L) in the vicinity of the MIP-1alpha gene, the locus surrounding the MIP-1alpha gene seems to be a "hot spring" that continuously produces new family genes. This evidence provides a new model, duplication and fusion, of the molecular basis for diversity within a gene family.     

A sequence-ready physical map of the region containing the human natural killer gene complex on chromosome 12p12.3-p13.2

We developed a sequence-ready physical map of a part of human chromosome 12p12.3-p13.2 where the natural killer gene complex (NKC) is located. The NKC includes a cluster of genes with structure similar to that of the Ca(2+)-dependent lectin superfamily of glycoproteins that are expressed on the surface of most natural killer (NK) cells and a subset of T cells. These killer cell lectin-like receptors (KLR) are involved in NK target cell recognition, leading to activation or inhibition of NK cell function. We used a number of sequence-tagged site (STS) markers from this region to screen two large insert bacterial artificial chromosome (BAC) libraries and a bacteriophage P1-derived (PAC) chromosome library. The clones were assembled into contiguous sets by STS content analysis. The 72-BAC and 11-PAC contig covers nearly 2 Mb of DNA and provides an average marker resolution of 26 kb. We have precisely localized 17 genes, 5 expressed sequence tags, and 49 STSs within this contig. Of this total number of STS, 30 are newly developed by clone-end sequencing. We established the order of the genes as tel-M6PR-MAFAL (HGMW-approved symbol KLRG1)-A2M-PZP-A2MP-NKRP1A (HGMW-approved symbol KLRB1)-CD69-AICL (HGMW-approved symbol CLECSF2)-KLRF1-OLR1-CD94 (HGMW-approved symbol KLRD1)-NKG2D (HGMW-approved symbol D12S2489E)-PGFL-NKG2F (HGMW-approved symbol KLRC4)-NKG2E (HGMW-approved symbol KLRC3)-NKG2A (HGMW-approved symbol KLRC1)-LY49L (HGMW-approved symbol KLRA1)-cen. This map would facilitate the cloning of new KLR genes and the complete sequencing of this region.     

Mapping of the Human Cysteine-Rich Intestinal Protein GeneCRIP1to the Human Chromosomal Segment 7q11.23

We report here on the mapping of a cDNA encoding for human cysteine-rich heart protein (HCRHP), a counterpart of the murine cysteine-rich intestinal protein CRIP. By somatic cell hybrid analysis and radiation hybrid mapping, we have located the geneCRIP1(HGMW-approved symbol) on the subcentromeric region of the q arm of human chromosome 7, flanking a deletion associated with Williams syndrome.     

Isolation and Characterization of a New Member of the HumanLy6Gene Family(LY6H)

TheLy6family of genes encodes glycosylphosphatidylinositol-anchored cell surface glycoproteins expressed on various types of cells. Intriguing patterns of expression ofLy6genes on specific subpopulations of lymphoid and myeloid cells suggest that Ly6 molecules may be involved in the development and homeostasis of hematopoietic cells. We have isolated a new member of the humanLy6gene family,LY6H,from a human fetal brain cDNA library. Fluorescencein situhybridization and radiation hybrid analyses assignedLY6Hto chromosome 8, where other members of theLy6gene family are also located. Northern analysis revealed thatLY6His highly expressed in particular subdivisions of human brain and also in MOLT-3 and -4 acute lymphoblastic leukemia cells. These data suggest that LY6H may play a role(s) in both the central nervous system and the immune system.     

Effects of Interactions BetweenPasteurella haemolyticaandBordetella parapertussisonin vitroPhagocytosis by Lung Macrophages

The aim of the work was to determine the effect of exposing ovine bronchoalveolar macrophages (BAM)in vivotoPasteurella haemolyticaand/orBordetella parapertussison the subsequent uptake and killing ofP. haemolyticaby these cellsin vitro. Exposurein vivotoP. haemolyticadid not affect the uptake ofP. haemolyticaby BAMin vitrobut reduced (P< 0·05) the intracellular killing of bacteria. Exposurein vivotoB. parapertussishad no significant effect on either the uptake of killing ofP. haemolytica in vitro. However, sequential exposurein vivotoB. parapertussisandP. haemolyticareduced both the ingestion (P< 0·05) and killing (P< 0·001) ofP. haemolytica in vitro. These results indicate that exposure toP. haemolyticacompromised the bacterial killing mechanisms of BAM and that synergy betweenB. parapertussisandP. haemolyticareduced the ability of BAM to ingest bacteria.     

Two new ectotroph-forming boletes from India

Pulveroboletus shoreae sp. nov. andXerocomus bakshii sp. nov. (Boletaceae) are described as new. Observations at the type locality show that they are both connected with short roots of the ectotrophic-mycorrhizal type, theXerocomus withPinus roxburghii, thePulveroboletus withShorea robusta, Fam. Dipterocarpaceae. This latter family has not been shown to form ectotrophs with ectotrophically mycorrhizal Basidiomycetes before our present research.