A comprehensive examination of protein sequences for evidence of internal gene duplication

Summary We have implemented a routine procedure for screening protein sequences for evidence of intragenic duplications. We tested 163 protein sequences representing 116 superfamilies of unrelated proteins. Twenty superfamilies contain proteins with internal gene duplications. The intragenic duplications detected can be divided into two major types. (1) One or more duplications of all or part of a gene produce a protein with two or several detectable regions of sequence homology. Sequences from 18 superfamilies contained this type of duplication. (2) Repeated reduplication of a small DNA segment can produce a protein that is repetitive over most of its length. Three superfamilies contain such repetitive sequences. We also investigated the limits of detection of ancient duplications using sequences derived by random mutation of a model sequence consisting of ten 10-residue repeats. The original repetitive nature of the sequence was usually detected after 250 point mutations even though the ancestral segment could not be accurately reconstructed.     

Paget's disease of bone in 14 British towns.

The radiological prevalence of Paget''s disease was studied in 14 towns. Routine radiographs showed that the disease was present in 5.4% of people aged 55 years and over. The disease was more prevalent in men than in women at all ages, and the prevalence increased with age. The three Lancashire towns studied (Preston, Bolton, and Blackburn) had higher rates than elsewhere. This probably reflects a real geographical variation in the prevalence of Paget''s disease in England and Wales.     

Methionyl-tRNA synthetase from Escherichia coli. Primary structure of the active crystallised tryptic fragment

A 3300-base segment of Escherichia coli chromosomal DNA, cloned into pBR322, will complement a methionine auxotroph in which the lesion is a defective methionyl-tRNA synthetase with a much reduced affinity for methionine. Crude extracts of these transformants contain elevated levels of a protein which has a subunit molecular weight of 66 000, methionyl-tRNA synthetase aminoacylation activity in vitro and which cross-reacts with anti-(methionyl-tRNA synthetase) antibodies. This polypeptide is very slightly larger than the well-characterised and crystallised tryptic fragment of methionyl-tRNA synthetase. A DNA sequence of 1750 residues at one end of the cloned insert codes for a non-terminated open reading frame in which we can locate a large number of methionyl-tRNA synthetase tryptic and chymotryptic peptides. We have also sequenced 300 nucleotides upstream of this coding segment where we find a large invert repeat in the putative methionyl-tRNA synthetase promoter region.     

Use of in situ hybridization to investigate the regulation of hippocampal corticosteroid receptors by monoamines.

The hippocampus receives major noradrenergic and serotoninergic (5-HT) innervations which interact with corticosteroid-sensitive cells. However, the subregional localization of these actions and the corticosteroid receptor types involved have not been defined and current ligand binding techniques for estimating corticosteroid receptors are hampered by several methodological limitations. We have developed in situ hybridization histochemical techniques to allow specific and sensitive estimation of glucocorticoid (GR) and mineralocorticoid receptor (MR) mRNA expression in rat hippocampus. Investigation of the effects of 5,7-dihydroxytryptamine lesions of 5-HT neurons showed significantly reduced GR and MR mRNA expression in some hippocampal subregions. Both abnormal 5-HT neurotransmission and excessive corticosteroid secretion are associated with major affective disorders, particularly depression. The crucial interaction between these two systems may occur, at least in part, at the level of regulation of hippocampal corticosteroid receptor expression.     

Relationship between Legionella pneumophila and Acanthamoeba polyphaga: physiological status and susceptibility to chemical inactivation.

Survival studies were conducted on Legionella pneumophila cells that had been grown intracellularly in Acanthamoeba polyphaga and then exposed to polyhexamethylene biguanide (PHMB), benzisothiazolone (BIT), and 5-chloro-N-methylisothiazolone (CMIT). Susceptibilities were also determined for L. pneumophila grown under iron-sufficient and iron-depleted conditions. BIT was relatively ineffective against cells grown under iron depletion; in contrast, iron-depleted conditions increased the susceptibilities of cells to PHMB and CMIT. The activities of all three biocides were greatly reduced against L. pneumophila grown in amoebae. PHMB (1 x MIC) gave 99.99% reductions in viability for cultures grown in broth within 6 h and no detectable survivors at 24 h but only 90 and 99.9% killing at 6 h and 24 h, respectively, for cells grown in amoebae. The antimicrobial properties of the three biocides against A. polyphaga were also determined. The majority of amoebae recovered from BIT treatment, but few, if any, survived CMIT treatment or exposure to PHMB. This study not only shows the profound effect that intra-amoebal growth has on the physiological status and antimicrobial susceptibility of L. pneumophila but also reveals PHMB to be a potential biocide for effective water treatment. In this respect, PHMB has significant activity, below its recommended use concentrations, against both the host amoeba and L. pneumophila.     

Determination of the cytokinin complement in healthy and witchesbroom malformed proteas

Cytokinins from normal and witchesbroom malformed stems of proteas were determined by radioimmunoassay following sample resolution by high-performance liquid chromatography (HPLC). Material from the early stages of shoot malformation had increased cytokinin concentrations which, over time, declined to the concentrations found in normal-growing stems. The cytokinin complement of the malformed structures was different from that of normal stems. The high concentrations of isopentenyladenosine detected appear to be related to the loss of the correlative inhibition of lateral buds and the development of the witchesbroom structures and may result from localized changes in cytokinin biosynthesis and/or metabolism.     

ON MISSING ENTRIES IN CLADISTIC ANALYSIS

Abstract The exact algorithms of two commonly used parsimony programs, Hennig86 by J. S. Farris and PAUP by D. Swofford, sometimes produce different solutions, and sometimes produce resolutions that are not supported by the data being analysed. The discrepancies apparently involve the treatment of missing entries, which can currently represent unknown data, inapplicable character and/or polymorphic taxa. Each of those potential sources of ambiguity is logically (if not computationally) different; with regard to binary characters, unknown data could be either 0 or 1, inapplicable characters are neither 0 nor 1 and polymorphisms are both 0 and 1. Resolutions that cannot be supported by any possible combination of known state attributions should either be flagged as such or suppressed entirely.     

Auxin controls Golgi-localized glucan synthetase activity in the maize mesocotyl

Decapitation or red light irradiation (R) inhibited growth and Golgi-localized glucan synthetase (GS I) activity in the mesocotyl of intact maize (Zea mays L.) seedlings. Applied auxin (indole-3-acetic acid) prevented the effects of R and of decapitation on both growth and GS I. Auxin applied several hours after irradiation prevented any further decline in GS I but did not restore it. Mesocotyl segments incubated in solution elongated in response to auxin but lost GS I with time regardless of the presence of exogenous auxin. An attached seed was necessary for maintenance of GS I in the dark-grown mesocotyl.Abbreviations GS glucan synthetase - IAA indole-3-acetic acid - R red light     

Photooxidation products of 7,12-dimethylbenz[a]anthracene.

The principal products of the photooxidation of 7,12-dimethylbenz[a]-anthracene (DMBA) in aqueous solutions by photooxidation induced by laboratory lighting have been characterized by high performance liquid chromatograms (HPLC), ultraviolet and mass spectrograms and by comparisons with authentic samples. The products identified were the 7,12-epidioxy-7,12-dihydro-7-12-dimethyl-, 7,12-dione, 7-hydroxymethyl-12-methyl-, 12-hydroxymethyl-7-methyl-, 7-formyl-12-methyl-, 12-formyl-7-methyl-, and 12-hydroxy-12-methyl-7-one derivatives of benz[a]-anthracene. The HPLC profile of products is similar to that obtained from oxidation of DMBA by 'one-electron' reagents, singlet oxygen, or liver microsomal metabolism. The first points of attack are the 7- and 12- positions. The mechanism of photooxidation appears to be generation of singlet oxygen by photodynamic effect of DMBA. None of the products is photosensitizing, however.