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Biomedical subjects

C P Gibbs

Publications and source records attributed to C P Gibbs.

At least 19 recordsLinked to original sources
PubMed1992

Interaction of two variable proteins (PilE and PilC) required for pilus-mediated adherence of Neisseria gonorrhoeae to human epithelial cells.

Pili confer the initial ability of Neisseria gonorrhoeae to bind to epithelial cells. Pilin (PilE), the major pilus subunit, and a minor protein termed PilC, reportedly essential for pilus biogenesis, undergo intra-strain phase and structural variation. We demonstrate here that at least two different adherence properties are associated with the gonococcal pili: one is specific for erythrocytes, which is virtually unaffected by PilE variation, and another is specific for epithelial cells, and is modulated in response to the variation of PilE. Based on this finding, mutants of a recA- strain were selected that had lost the ability to bind to human cornea epithelial cells (A-) but retained the ability to form pili (P+) and to agglutinate human erythrocytes (H+). The adherence-negative mutants failed to produce detectable levels of PilC1 or PilC2 proteins, representing piIC phase variants generated in the absence of RecA. The A- pilC phase variants were indistinguishable from their A+ parents and spontaneous A+ revertants with regard to the amount of PilE produced and its electrophoretic mobility, the degrees of piliation and haemagglutination, and the pilE nucleotide sequence. These data demonstrate a central role for PilC in pilus-mediated adherence of N. gonorrhoeae to human epithelial cells and further indicate that neither PilC1 nor PilC2 is obligatory for the assembly of gonococcal pili.

Amino Acid Sequence↗
PubMed1991

The opacity proteins of Neisseria gonorrhoeae strain MS11 are encoded by a family of 11 complete genes.

Variants of Neisseria gonorrhoeae MS11 show distinct colony morphologies because of the expression of a class of surface components called opacity (Opa, PII) proteins. Southern analyses combined with molecular cloning of genomic DNA from a single variant of MS11 has identified 11 opa genes contained in separate loci. These opa genes code for distinct opacity proteins which are distinguishable at their variable domains. The opa gene analyses were also extended to divergent variants of MS11. These studies have shown that, during in vitro and in vivo culture, 10 of the 11 opa genes did not undergo significant change in their primary sequence. However, in these variants, one gene (opaE) underwent non-reciprocal inter-opa recombinations to generate newer Opa variants. Phylogenic analysis of the opa gene sequences suggests that the opa gene family have evolved by a combination of gene duplication, gene replacement and partial inter-opa recombination events.

Amino Acid Sequence↗
PubMed1991

Physical and genetic map of the Neisseria gonorrhoeae strain MS11-N198 chromosome.

A macro-restriction map of the Neisseria gonorrhoeae chromosome was constructed using the enzymes Nhel and Spel. Combinations of one- and two-dimensional electrophoresis of completely or partially digested chromosomal DNA were performed to align the restriction fragments. The chromosome is circular, with an estimated size of 2.33 Mb +/- 35 kb. A genetic map was derived from the physical map; positions of over 60 defined loci were determined by Southern hybridization.

Blotting, Southern↗
PubMed1989

Reassortment of pilin genes in Neisseria gonorrhoeae occurs by two distinct mechanisms.

Phase and antigenic variation of pilin expression in Neisseria gonorrhoeae result from recombination events in which variant sequences from one of the silent loci (pilS) are transferred to the expression locus (pilE). Such rearrangements were originally thought to be gene conversions, but findings showing that phase variation is partially inhibited by DNase I, that piliated (P+) cells are highly competent for DNA uptake and that gonococci readily undergo autolysis in culture, led to the suggestion that pilin variation occurs through transformation by exogenous DNA. We have developed a simple method for the selection of non-piliated (P-) cells and have evaluated naturally occurring P+ to P- transitions. Two primary pathways of pilin variation can be distinguished--transformation-mediated recombination, which is influenced by culture conditions and inhibited by DNase I, and intragenomic reciprocal recombination, which is unaffected by DNase I. Furthermore, we demonstrate that both piliated and revertible P- cells are competent for DNA uptake, an essential prerequisite of the first pathway.

Bacterial Outer Membrane Proteins↗
PubMed1988

Cloning and transcriptional analysis of a variant surface glycoprotein gene expression site in Trypanosoma brucei.

The variant surface glycoprotein (VSG) gene expression site in Trypanosoma brucei variant 117a has been mapped to a point about 40 kb upstream from the VSG gene. Sequences upstream from the previously identified [Cully, D.F. et al. (1985) Cell 42, 173-182] expression site associated gene (ESAG-I) have been cloned and a stable 1.3 kb transcript has been localized immediately 5' to ESAG-I. This transcript is in the same orientation and approximately as abundant as the ESAG-I message. A highly conserved region, of which at least 15 copies are present in the genome, has been identified further upstream. A stable transcript corresponding to this region was not detected in variant 117a, but a 1.7 kb transcript was detected in variant 221a. In isolated nuclei, representative sequences from the 117a, expression site were transcribed unidirectionally at similar rates, and transcription was insensitive to alpha-amanitin.

Animals↗
PubMed1987

DNA sequence encoding the amino-terminal region of the human c-src protein: implications of sequence divergence among src-type kinase oncogenes.

We sequenced the 5'-coding region of the human c-src gene, exons 2 through 5, corresponding to one-third of the human c-src protein consisting of 536 amino acids. Sequence analysis of the src type of protein kinases revealed that the amino-terminal region encoded by exon 2 contains sequences specific for the src proteins and raised the possibility that this region is involved in the recognition of a src-specific substrate(s) or receptor(s).

Amino Acid Sequence↗
PubMed1986

Identification of proteins encoded by variant surface glycoprotein expression site-associated genes in Trypanosoma brucei.

The variant surface glycoprotein (VSG) genes of Trypanosoma brucei may be transcribed from several distinct telomeric expression sites (ESs). The mechanism responsible for regulating potential expression sites is unknown. Two members of a pleomorphic family of expression site associated genes (ESAGs) have been cloned and sequenced. By examination of the DNA sequences we inferred that ESAGs encode amphiphilic glycoproteins. Fragments of two ESAGs were inserted into the Escherichia coli expression vectors pATH and pEX. Antisera to the resulting anthranilate synthetase ESAG protein (ESAGP) fusion protein immune precipitated a 46 kDa glycoprotein from detergent extracts of T. brucei. In the presence of tunicamycin, the size of the immune-precipitated protein was reduced to 36 kDa, corresponding to the molecular weight predicted by the ESAG sequence. The 36 kDa and 46 kDa proteins were absent from procyclic culture forms of T. brucei.

Animals↗
PubMed1986

Obstetric anesthesia: a national survey.

To assess obstetric anesthesia in the United States, and to determine why more anesthesia personnel are not involved in this subspecialty, a questionnaire was sent to the heads of obstetric and anesthesia services in 1,200 hospitals. Both obstetric and anesthesia respondents agreed on several characteristics of obstetric anesthesia that inhibit more participation by anesthesia personnel. Among others, they identified that: the unpredictability of labor and delivery makes scheduling difficult; obstetricians tend to dictate type and timing of anesthesia; the risk of malpractice claims is increased for obstetric anesthesia; and, finally, larger obstetric services would make it more practical to provide anesthesia services. Regarding availability of personnel and procedures, obstetric units with less than 500 deliveries per year were considerably more under-staffed than the larger units in most areas studied. When general anesthesia was used for cesarean section in these units, it was provided by, or given under the direction of, an anesthesiologist only 44% of the time, whereas in the hospitals with more than 1,500 deliveries per year, an anesthesiologist was present 86% of the time. Likewise, in the small units, personnel classified as "others" were responsible for newborn resuscitation in 24% and 43% of instances after cesarean section and vaginal delivery, respectively. In the hospitals with more than 1,500 deliveries, comparable figures were 4% and 2%, respectively.

Anesthesia, Epidural↗
PubMed1985

Isolation and structural mapping of a human c-src gene homologous to the transforming gene (v-src) of Rous sarcoma virus.

We have utilized a lambda Charon 4A human genomic library to isolate recombinant clones harboring a highly conserved c-src locus containing nucleotide sequences homologous to the transforming gene of Rous sarcoma virus (v-src). Four overlapping clones spanning 24 kilobases of cellular DNA were analyzed by restriction endonuclease mapping. Human c-src sequences homologous to the entire v-src region are present in a 20-kilobase region that contains 11 exons as determined by restriction mapping studies utilizing hybridization to labeled DNA probes representing various subregions of the v-src gene and by preliminary DNA sequencing analyses. A considerable degree of similarity exists between the organization of the human c-src gene and that of the corresponding chicken c-src gene with respect to exon size and number. However, the human c-src locus is larger than the corresponding chicken c-src locus, because many human c-src introns are larger than those of chicken c-src. alu family repetitive sequences are present within several human c-src introns. This locus represents a highly conserved human c-src locus that is detectable in human cellular DNAs from various sources including placenta, HeLa cells, and WI-38 cells.

Avian Sarcoma Viruses↗
PubMed1985

Human cellular src gene: nucleotide sequence and derived amino acid sequence of the region coding for the carboxy-terminal two-thirds of pp60c-src.

The nucleotide sequence of the 3' two-thirds of a highly conserved, molecularly cloned human cellular src gene (c-src) has been determined. This region of the c-src gene encodes the tyrosine kinase domain of the cellular src protein (pp60c-src) and corresponds to exons 6 through 12 of the chicken c-src gene, as well as nucleotides 545 to 1542 of the Rous sarcoma virus src gene (v-src). The human c-src sequence is very strongly conserved with respect to both the chicken c-src and the Rous sarcoma virus v-src genes, with nearly 90% nucleotide homology observed in this region. Amino acid sequence conservation in this region is even greater; 98% of the amino acids are conserved between human and chicken c-src. Furthermore, the exon sizes and the locations of the exon-intron boundaries are identical in the human and chicken c-src genes. However, sequences within the introns have not been conserved, and the introns within the human c-src gene are significantly larger than the corresponding introns within the chicken c-src gene. The strong amino acid conservation between the carboxy-terminal two-thirds of pp60c-src of species as divergent as humans and chickens suggests that this portion of the pp60c-src protein specifies one or more functional domains that are of great importance to some aspect of normal cellular growth or differentiation.

Amino Acid Sequence↗
PubMed1984

Extensive homology exists between Marek disease herpesvirus and its vaccine virus, herpesvirus of turkeys.

Marek disease is a lymphomatous disease of chickens caused by infection of a herpesvirus, Marek disease virus (MDV). Marek disease is the only neoplastic disease for which a successful vaccine has been developed. The vaccine virus, herpesvirus of turkeys (HVT), is non-oncogenic in chickens. Despite the strong antigenic relationship between these viruses, previous studies showed that the two viral DNAs share little or no homology. Using less stringent hybridization conditions and methods that greatly improve the reassociation kinetics, we have reexamined the sequence homology between MDV and HVT DNA. We report here that HVT and MDV are far more closely related than previously reported. The homology between these two viral DNAs ranges between 70% and 80% at the nucleotide level and appears to extend over 90-95% of the respective genomes. Under the low stringency conditions used, MDV DNA fails to cross-hybridize with DNA from feline rhinotracheitis virus, an antigenically unrelated herpesvirus with a G-C content identical to that of MDV and HVT.

Base Sequence↗