Mammalian Genome

Ibaraki, K.; Kozak, C.; Wewer, U.; Albrechtsen, R.; Young, M.

doi: 10.1007/BF00354289pmid: 8563165

Tetranectin is a plasminogen-binding tetrameric protein originally isolated from plasma. Expression of tetranectin appears ubiquitous, although particularly high expression is noted in the stroma of malignant tumors and during mineralization. To dissect the molecular basis of tetranectin gene regulation, mouse tetranectin cDNA was cloned from a 16-day-old mouse embryo library. Sequence analysis revealed a 992-bp cDNA with an open reading frame of 606 bp, which is identical in length to the human tetranectin cDNA. The deduced amino acid sequence showed high homology to the human cDNA with 76% identity and 87% similarity at the amino acid level. Sequence comparisons between mouse and human tetranectin and some C-type lectins confirmed a complete conservation in the position of six cysteines as well as numerous other amino acid residues, indicating an essential structure for potential function(s) of tetranectin. The sequence analysis revealed a difference in both sequence and size of the noncoding regions between mouse and human cDNAs. Northern analysis of the various tissues from mouse, rat, and cow showed the major transcript(s) to be approximately 1 kb, which is similar in size to that observed in human. Although additional minor bands of 1.5 and 3.3 kb were found in Northern blots, RT-PCR (reverse transcription polymerase chain reaction) analysis failed to provide evidence that these minor bands are products of the tetranectin gene. Finally, the genetic map location for this gene, Tna, was determined to be on distal mouse Chromosome (Chr) 9 by analysis of two sets of multilocus crosses.

Duncan, M.; Lieman, J.; Chada, K.

doi: 10.1007/BF00354290pmid: 8563166

The autosomal recessive mouse mutation, germ cell dificient, gcd, manifests as infertility in both sexes owing to improper migration and/or proliferation of primordial germ cells during embryonic development. Mice harboring this mutation have been hypothesized to be animal models of the human syndromes, premature ovarian failure and Sertoli cell only syndrome. Since the gcd mutation arose from the insertion of over 100 kb of foreign DNA into the chromosome during a transgenic mouse experiment, fluorescent in situ hybridization with the transgene as a probe was used to determine the chromosomal position of the gcd locus. DAPI chromosomal banding in conjunction with double labeling with the α1(I) collagen gene revealed that the gcd locus is situated on mouse Chromosome (Chr) 11A2–3. Two candidate genes, Lif and Oncostatin M, map near the gcd locus; however, Southern blot hybridization analysis revealed no gross rearrangements in these genes in gcd mice. The chromosomal position of the gcd locus will prove valuable in the search for other candidate genes as well as a landmark for positional cloning experiments.

Mullick, A.; Groulx, N.; Trasler, D.; Gros, P.

doi: 10.1007/BF00354291pmid: 8563167

Looptail (Lp) is a mutation on the distal portion of mouse Chromosome (Chr) 1 that affects neurulation in mouse and is phenotypically expressed by appearance of an open neural tube along the entire antero-posterior axis of the embryo (craniorachischisis). Nhlh1, a member of the basic helix-loop-helix family of transcription factors, is expressed in the developing neural tube in structures affected by the Lp mutation and has been regionally assigned to the distal part of mouse Chr 1. Using a large panel of looptail animals from an (Lp/+ x SWR/J)F1 x SWR/J segregating backcross progeny, we have determined that Nhlh1 maps very close to Lp, with no recombinant detected in 500 informative animals tested; both map within a 0.6-cM segment defined as D1Mit113/apoa2/Fcer1γ-(0.4 cM)-Nhlh1/Lp-(0.2 cM)-Fcer1α/D1Mit149/Spna1. Nucleotide sequencing of Nhlh1 cDNA clones from wild type (WT) and Lp/Lp embryos failed to identify sequence alterations associated with the mutant phenotype. Southern hybridization of genomic DNA from WT and Lp/Lp embryos failed to identify specific rearrangements at or near the Nhlh1 locus, and Northern RNA blotting and RT-PCR evaluation of Nhlh1 mRNA expression indicated that both the levels and types of Nhlh1 mRNAs produced in WT and Lp/Lp embryos were indistinguishable. These studies suggest that Nhlh1 and Lp are not allelic. Nevertheless, Nhlh1 is the Chr 1 marker most tightly linked to Lp identified to date and can, therefore, be used as an excellent entry probe to clone the Lp region.

Khlebodarova, T.; Malchenko, S.; Matveeva, N.; Pack, S.; Sokolova, O.; Alabiev, B.; Belousov, E.; Peremislov, V.; Nayakshin, A.; Brusgaard, K.; Serov, O.

doi: 10.1007/BF00354292pmid:

Schmutz, S.; Marquess, F.; Berryere, T.; Moker, J.

doi: 10.1007/BF00354293pmid: 8563169

Five Charolais families known to segregate for both horned and polled were selected and tested for linkage analysis by use of microsatellites and karyotyping for Robertsonian translocation 1;29. No recombinants were found between any of these markers and the polled phenotype or each other. When statistical analysis was performed, the logarithm of the odds (LOD) indicated that there was 100% linkage occurring between the markers and the phenotype (p<0.001). These microsatellite markers, TGLA49 and BM6438, can be assumed to be very close to the actual gene that determines the polled phenotype. Another linked marker, SOD1, was physically mapped, which places all of these markers within 1q12–14, very near the centromere of Chromosome (Chr) 1. A homozygous polled cow was identified in this study by following the alleles at both markers and the phenotypes in her family.

Stone, R.; Pulido, J.; Duyk, G.; Kappes, S.; Keele, J.; Beattie, C.

doi: 10.1007/BF00354294pmid: 8563170

A bovine genomic phagemid library was constructed with randomly sheared DNA. Enrichment of this single-stranded DNA library with CA or GT primers resulted in 45% positive clones. The 14% of positive clones with (CA · GT)>12, and not containing flanking repetitive elements, were sequenced, and the efficiency of marker production was compared with random M13 bacteriophage libraries. Primer sequences and genotyping information are presented for 390 informative bovine microsatellite markers. The genomic frequency for 11 tri- and tetranucleotide repeats was estimated by hybridization to a lambda genomic library. Only GCT, GGT, and GGAT were estimated to have a frequency of >100 per genome. Enrichment of the phagemid library for these repeats failed to provide a viable source of microsatellite markers in the bovine. Comparison of map interval lengths between 100 markers from the enriched library prepared from randomly sheared DNA and M13 bacteriophage libraries prepared from Mbo1 restriction digests suggested no bias in skeletal genomic coverage based on source of small insert DNA. In conclusion, enrichment of the bovine phagemid library provides a sufficient source of microsatellites so that small repeat lengths and flanking repetitive sequences common in the bovine can be eliminated, resulting in a high percentage of informative markers.

Robinson, P.; Leek, J.; Thompson, J.; Carr, I.; Bailey, A.; Moynihan, T.; Coletta, P.; Lench, N.; Markham, A.

doi: 10.1007/BF00354295pmid: 8563171

We have identified a novel ubiquitin conjugating enzyme gene, L-UBC, which maps to human Chromosome (Chr) 14q24.3. This is also the location of the major early onset familial Alzheimer's disease gene (FAD3). L-UBC encodes a protein that demonstrates homology to the yeast ubiquitin conjugating enzyme, UBC-4, and human UbcH5. Their functions are to ubiquitinate specific proteins targeted for degradation. The protein also exhibits very strong homology to a rabbit protein, E2-F1, which mediates p53 degradation driven by papilloma virus E6 protein in vitro. The accumulation of specific proteins that have undergone aberrant processing in neurofibrillary tangles and amyloid plaques is the classic pathological feature in brains of Alzheimer's disease patients. Abnormal ubiquitination has previously been suggested to play a role in the etiology of Alzheimer's disease. This gene therefore represents a plausible candidate gene for FAD3.

Goddard, A.; Yuan, J.; Fairbairn, L.; Dexter, M.; Borrow, J.; Kozak, C.; Solomon, E.

doi: 10.1007/BF00354296pmid: 8563172

PML, a Ring-finger protein, participates in the disruption of normal myeloid differentiation when fused to the retinoic acid receptor alpha (RARα) by the translocation between Chromosomes (Chrs) 15 and 17 in acute promyelocytic leukemia (APL). As an initial step in the characterization of PML in species other than human, a murine cDNA clone of the PML gene was isolated and sequenced, and the intron/exon organization of the murine locus determined. The predicted amino acid sequence of the mouse PML protein shows 80% similarity to that of its human homolog. However, the mouse and human proteins show greater than 90% similarity in the proposed functional domains of the proteins. Despite its role in the etiology of APL, PML expression is not detectably altered during granulocytic differentiation in a murine in vitro system. Chromosomal localization of the Pml locus by somatic cell hybrids and by linkage analysis indicates that the gene maps to a region of mouse Chr 9 with known linkage homology to the region on human Chr 15q to which PML has been localized.

Davies, P.; Melanitou, E.; Asano, M.; Avner, P.; Montagutelli, X.

doi: 10.1007/BF00354297pmid: 8563173

White, R.; Angeloni, S.; Pasztor, L.

doi: 10.1007/BF00354298pmid: 8563174