Unequal synthesis and differential degradation of propionyl CoA carboxylase subunits in cells from normal and propionic acidemia patients.

We have characterized further the molecular basis of human inherited propionyl CoA carboxylase deficiency by measuring steady state levels of the mRNAs coding for the enzyme's two protein subunits (alpha and beta) and by estimating initial synthesis and steady state levels of the protein subunits in skin fibroblasts from controls and affected patients. We studied cell lines from both major complementation groups (pccA and pccBC) corresponding, respectively, to defects in the carboxylase's alpha and beta subunits. Analysis of pccA lines revealed the absence of alpha chain mRNA in three and an abnormally small alpha-mRNA in a fourth. Despite the presence of normal beta-mRNA in each of these pccA lines, there was complete absence of both alpha and beta protein subunits under steady state conditions, even though new synthesis and mitochondrial import of beta precursors was normal. Results in nine pccBC lines revealed normal alpha mRNA in each, while the amounts of beta-mRNA were distinctly reduced in every case. Correspondingly, alpha protein subunits were present in normal amounts at steady-state, but beta subunits were uniformly decreased. In addition, in six of the nine beta deficient cell lines, partially degraded beta-subunits were observed. To help interpret these results, synthesis and stability of carboxylase subunits were studied in intact HeLa cells using a pulse-chase protocol. Whereas alpha chains were stable over the four hour interval studied, beta chains--initially synthesized in large excess over alpha chains--were degraded rapidly reaching equivalence with alpha chains after two hours.(ABSTRACT TRUNCATED AT 250 WORDS)     

The alpha-A-crystallin and cystathionine beta-synthase genes are physically very closely linked in proximal mouse chromosome 17

Murine genes homologous to those contributing to the Down syndrome (DS) phenotype in man are currently of interest because of their potential for providing animal models for the study of specific DS symptoms. Most of the genes mapping to human chromosome 21q22, where the DS genes are concentrated, are related to sequences located on mouse chromosome 16. Others, however, are known to map to mouse chromosome 10, and two genes, cystathionine beta-synthase (Cbs) and alpha-A-crystallin (Crya-1), have been localized to the proximal portion of mouse chromosome 17. In this paper, we show that the two genes mapping to human chromosome 21q22 and mouse chromosome 17 are very tightly linked in mouse, being separated by at least 70 kb, but not more than 130 kb. The very close physical linkage of mouse Cbs and Crya-1, combined with data that localize homologs of the closely flanking markers H2k and Pim-1 to human chromosome 6, suggests that the human 21q22/mouse chromosome 17 conserved segment is of a very limited total physical size and is likely to contain a relatively small number of genes.     

The "suspicious" gynecologic smear

"Suspicious" gynecologic smears from 842 patients over a seven-year period were analyzed for their causes and outcomes. The frequency of the cytologic diagnosis of "suspicious" ranged between 0.5% in 1979 and 1.44% in 1975 of all smears examined. Review of the smears showed that this classification was used to report a variety of conditions, including equivocal possible precancerous changes as well as the presence of severe inflammation, degenerative or atrophic changes, abnormal glandular cells and metaplasia. The cytologic follow-up, following anti-inflammatory or hormonal therapy, showed a conversion to negative findings in 65.1% of all cases, usually within 12 months. In 294 cases, histologic analysis became necessary, revealing precancerous changes or cancer in 147 patients (17.5% of the study group). Smears of postmenopausal women with suspicious glandular or endometrial cells received special analysis. Significant numbers of such cases had histologic findings positive for malignancy (20% of smears with glandular cells and 21.3% with endometrial cells), as did also smears showing post-irradiative changes (34.6%) or atrophic and degenerative changes (17.1%). Therefore, "suspicious" smears in these groups were considered to indicate an increased risk of malignancy. A regimen for the proper management of cases with "suspicious" smears has been established.     

Composition of Caulobacter crescentus murein synthesized in the presence of glycine

Abstract Glycine added to the growth medium of Caulobacter crescentus was found to substitute Cterminal alanine in the peptide side chains of the murein of this species. Murein synthesized in vivo and in vitro in the presence of glycerine was poorly crosslinked as was new murein formed in the presence of the amino acid. The reduced cross-linkage seems to be due to the effect of glycine on the formation of trimeric muropeptides as revealed by high-performance liquid chromatography (HPLC) muropeptide analysis of murein formed in the presence and absence of the amino acid.     

Are there insertions in the ribosomal DNA of vertebrates?

We have analysed the Eco RI restriction pattern of rDNA of the newt Triturus vulgaris and of some other amphibian species by Southern blotting and hybridization with nick-translated Xenopus rDNA prepared from the recombinant plasmids pXlr11 and pXlr12 (21). After hybridization with r11, the 28S coding fragments become visible in two bands, a prominent one of 5.3 kb and a weak band of 5.9 kb representing about 8% of the 28S genes. The evidence obtained so far by additional digestions with Bam HI and Bgl II indicates that in this species and in Triturus helveticus the coding regions of the 5.9 kb fragments are interrupted by an insertion 0.6 kb in length located in a 1.6 kb Bgl II fragment at the 3' end of the Eco RI fragment, which we believe to be the first described in a vertebrate.     

Localization of the human erbB-2 gene on normal and rearranged chromosomes 17 to bands q12-21.32

Through the use of a cDNA probe, the human erbB-2 gene was localized by in situ hybridization of normal human chromosomes at 17q11-q21. In situ hybridization of chromosomes derived from fibroblasts carrying a constitutional 15;17t(q22.3;q11.21) translocation showed that the erbB-2 gene was relocated on the rearranged chromosome 15. These results as well as grain localization on prophase chromosomes locate the erbB-2 gene at 17q12-q21.32. This localization may facilitate the search for human malignancies with chromosome changes involving the erbB-2 gene.     

Is male back space limiting? An investigation into the reproductive demography of the giant water bug,Abedus indentatus (Heteroptera: Belostomatidae)

Because male giant water bugs in the subfamily Belostomatinae provide parental care by brooding eggs on their back, an accurate assessment can be made of both the actual and potential reproductive capacity of males. Two operational sex-ratio (OSR) indices were developed and empirically measured for a population of giant water bug, Abedus indentatus,in California. One index was based on reproductive rates measured in the laboratory; the other index was based on reproductive resources observed in the field. Both OSR indices suggest that the operational sex ratio fluctuates between maleskewed ratios in the summer and femaleskewed ratios in the winter. This pattern appears to be the consequence of two factors. First, the adult sex ratio is significantly female biased. Second, although males can outreproduce females at high ambient temperatures, the reverse is true at low temperatures. Possible reasons for the female-skewed adult sex ratio are examined, including differential recruitment, differential mortality, and sampling bias.