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[Effect of diphenylhydantoin on a developed epileptogenic focus in cats with split cerebral hemispheres].

The purpose of this work was to assess the effects of DPH on a developed epileptogenic focus in cats with split cerebral hemispheres. The investigations were carried out on 12 cats with a chronic epileptogenic focus produced by means of aluminum method in the right motor area. In all cats the epileptogenic focus was found in EEG. All animals received DPH in daily doses of 8-15 mg/kg. In 2 cats they appeared before beginning of treatment. One of these cats died after 3 days from status epilepticus, the other survived status epilepticus and died after 42 days of DPH administration with signs of intoxication. In 3 cats clinical seizures developed during DPH treatment after 30.84 and 210 days. DPH was given during from 171 to 314 days. Clinical seizures appeared in these cats only sporadically and the animals were sacrificed after completion of investigations. In 7 out of 12 cats clinical seizures failed to develop despite presence of bioelectrically active epileptogenic seizures in the right motor area. Administration of DPH in cats with developed epileptogenic focus failed to prevent clinical seizures. In cats with seizures their control was limited by drug toxicity. In all animals toxic effects were observed although the serum DPH level was in the range 8-20 mug/ml.

Aluminum↗

Feline oncornavirus-associated cell membrane antigen. II. Antibody titers in healthy cats from household and laboratory colony environments.

Antibody titers to the feline oncornavirusassociated cell membrane antigen (FOCMA) were determined for 447 healthy cats from laboratory colony and household environments. Only 2.7 percent of 221 cats from colony environments were antibody positive as compared to 50.4 percent of 256 cats from household environments. Incidence of FOCMA antibody and geometric mean antibody titer for pet cats from New York City representing single cat apartment habitats were substantially lower than values for unscreened cats from the Boston, Glasgow, and Detroit suburban environments. Geometric mean antibody titer for young adults in the Boston population was significantly higher than titers for kittens or aged cats. This may be due to greater mobility resulting in increased exposure to other cats. In contrast to the high frequency of FOCMA antibody positivity in pet-cat populations, less than 2 percent of the same groups were positive for virus group-specific antigen in peripheral blood leukocytes and platelets. This was interpreted as an indication that many more cats became infected with feline leukemia virus under natural conditions that the number developing persistent virus infection and/or clinical leukemia.

Animals↗

Effect of recombinant human granulocyte colony-stimulating factor on hematopoiesis in normal cats.

The objective of this study was to determine how recombinant human granulocyte colony-stimulating factor (rhG-CSF) affects hematopoiesis in normal cats. Recombinant human G-CSF was given at 3.0, 5.0, and 10.0 micrograms/kg to two cats each s.c. twice daily for 21 days. This resulted in significant (p less than 0.01) elevations of peripheral blood neutrophils from 3.0- to 9.2-fold above pretreatment levels and significantly (p less than 0.02) above levels of nontreated control cats (n = 4). A statistically significant dose-related response was not seen at these dosages in any parameter evaluated. The period of maximum neutrophilia occurred between days 10 and 14 of rhG-CSF treatment, with maximum neutrophil counts ranging from 20,370 cells/microliters to 61,400 cells/microliters (normal is less than 12,500). Lymphocytosis (greater than 7000 lymphocytes/microliters) and monocytosis (greater than 850 monocytes/microliters) were observed in 50% of the cats receiving rhG-CSF during the period of maximal neutrophil stimulation. Monocyte counts in treated cats were significantly (p less than 0.01) elevated over those of treatment controls on days 12-17. Lymphocyte numbers in rhG-CSF-treated cats were significantly elevated (p less than 0.05) over pretreatment controls on days 12 and 14 of rhG-CSF treatment. No significant changes were observed in reticulocyte counts, platelet counts, or hematocrit levels. By day 19, neutrophil levels had dropped significantly (p less than 0.01) from the maximum neutrophil levels, with one cat attaining a normal blood neutrophil count by day 21 of rhG-CSF treatment. Marrow aspirates revealed an overall increase in marrow cellularity through day 14 of treatment in rhG-CSF-treated cats, with increased myeloid:erythroid ratios (two- to ninefold) over those of nontreated controls. The erythroid and lymphoid component of the marrow decreased from day 0 to day 14, whereas the early myeloid progenitors (myeloblasts, progranulocytes, and myelocytes) increased significantly (p less than 0.05). No significant differences in the percentage of later myeloid forms in the marrow were observed over the treatment period. In vitro colony-forming assays of marrow obtained from treated cats revealed increases in granulocyte-macrophage colony-forming units (CFU-GM) through day 14, with subsequent decreases by day 21 of rhG-CSF treatment. Recombinant human G-CSF was also effective at in vitro stimulation of feline marrow cells from untreated cats in a dilution study, with maximal CFU-GM formation at 0.1 microgram rhG-CSF/ml assay.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Effect of sodium bicarbonate infusion on serum osmolality, electrolyte concentrations, and blood gas tensions in cats.

The effects of single IV injections of sodium bicarbonate (0.5 mEq/kg of body weight, 1 mEq/kg, 2 mEq/kg, and 4 mEq/kg) on serum osmolality, serum sodium, chloride, and potassium concentrations, and venous blood gas tensions in 6 healthy cats were monitored for 180 minutes. Serum osmolality increased and remained significantly (P less than 0.05) increased for 120 minutes in cats given 4 mEq of sodium bicarbonate/kg. Serum sodium was increased significantly (P less than 0.05) for 30 minutes in cats given 4 mEq of sodium bicarbonate/kg. Serum sodium decreased and remained significantly (P less than 0.05) decreased for 120 minutes in cats given 1 g of 20% mannitol/kg, and serum osmolality was significantly (P less than 0.05) decreased at 30 and 60 minutes. Serum chloride decreased significantly (P less than 0.05) for 10 minutes in cats given 1 mEq of sodium bicarbonate/kg, and was significantly decreased for 30 minutes in cats given 2 mEq and 4 mEq of sodium bicarbonate/kg. Serum chloride decreased and remained significantly (P less than 0.05) decreased for 30 minutes in cats given 1 g of 20% mannitol/kg. Serum sodium and serum osmolality did not change significantly (P less than 0.05) in cats given 4 ml of 0.9% sodium chloride/kg. Serum potassium decreased significantly (P less than 0.05) for 10 minutes in cats given 1 mEq of sodium bicarbonate/kg, and for 120 minutes in cats given 2 mEq/kg or 4 mEq/kg. There was a significantly (P less than 0.05) greater decrease in serum potassium that lasted for 30 minutes after given sodium bicarbonate at the dosage of 4 mEq/kg, compared with other dosages given.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Interleukin-2 suppression in cats naturally infected with feline leukemia virus.

Feline peripheral blood lymphocytes (PBLs) produce interleukin-2 (IL-2) when stimulated with concanavalin A (Con A) or the combination of the Ca2+ ionophore A23187 (A2) plus the phorbol ester phorbol-12-myristate-13-acetate (PMA). To determine the status of T helper cell function in cats naturally infected with feline leukemia virus (FeLV), PBLs from 19 normal cats and 49 FeLV-infected (34 asymptomatic and 15 symptomatic) cats were tested for their ability to produce IL-2 in response to Con A or A2/PMA stimulation. In the case of Con A stimulation, 21, 41, and 67% of normal, FeLV-infected asymptomatic, and FeLV-infected symptomatic cats, respectively, failed to produce detectable levels of IL-2. In contrast, A2/PMA stimulation yielded high titers of IL-2 in 100% of the normal cats, 92% of asymptomatic cats, but only 44% of clinically ill cats. All of the A2/PMA nonresponders in the FeLV-infected symptomatic population were also unresponsive to Con A. T Helper immunosuppression was observed in cats with FeLV-induced lymphoproliferative diseases as well as cats with FeLV-associated acquired immune deficiency syndrome-like diseases. These data suggest that TH immunosuppression in naturally infected viremic cats may precede development of clinical disease. Moreover, the ability to restore TH function with A2/PMA is diminished with development of clinical disease.

Animals↗

Response to intradermal skin testing with four cat allergen preparations in healthy and allergic dogs.

Intradermal skin testing with 4 cat allergens was performed on 40 dogs. The allergens used were a commercial preparation, cat allergen 1, cat pelt extract, and cat serum. Twenty dogs had inhalant allergies (canine atopy) and 20 dogs were healthy. The dogs in these 2 groups were further allotted to groups of dogs with or without exposure to cats. Cat pelt extract was the only allergen that caused a statistically significant (P less than 0.025) difference in the number of positive reactions in healthy vs allergic dogs, with healthy dogs having the greater number of positive reactions. Seven (35%) of the 20 dogs with no known exposure to cats had positive reactions to at least one of the allergens. In dogs that had been exposed to cats, positive reactions developed in twice the number of dogs without clinical signs of canine atopy, compared with those with clinical signs of canine atopy. These data showed that dogs do become sensitized to cat allergen, but that this sensitization may not indicate known exposure to cats or clinical disease.

Allergens↗

Discrimination learning of the vertical and horizontal light spot oscillations in normal and visually deprived cats.

Three groups of cats were used: cats deprived binocularly of patterned vision from birth (BD cats), control cats reared in the laboratory with opened eyes (C cats) and cats reared in a rural environment during the first months of life (N cats). The cats were trained to discriminate vertical vs. horizontal oscillations of a light spot for food reward. The task was difficult for all cats. The presumable reason was that the majority of neurones discriminates best between two opposite directions of movement and this mechanism was useless in the task. The learning was only slightly impaired in the BD group and only as compared with the N group. Our main conclusion is that a reduced number of directionally selective neurons in BD cats was still sufficient for movement discrimination learning.

Animals↗

Preclinical evaluation of a sterically stabilized liposome-encapsulated cisplatin in clinically normal cats.

OBJECTIVE: To evaluate the effects of administration of a sterically stabilized liposome-encapsulated cisplatin (SSL-CDDP) to cats. ANIMALS: 4 clinically normal cats. PROCEDURE: 2 of the cats were given multiple i.v. injections of SSL-CDDP at a dosage of 70 mg of free CDDP equivalent/m2 of body surface area at 3-week intervals. The other 2 cats received single i.v. injections of identical liposome preparations not containing CDDP. Vital signs; appetite; attitude; hematologic, serum biochemical, and urinalysis findings; and thoracic radiographic views were evaluated at predetermined intervals. RESULTS: Sterically stabilized liposome-encapsulated cisplatin was well tolerated by all cats. The only significant alterations in measured variables were an increase in serum cholesterol concentration 2 days after injection, and repeatable pyrexia in the cats receiving SSL-CDDP that began 10 to 12 hours after injection and continued for 18 hours, peaking at 40.5 to 41 C. Alterations in rectal temperature were not significant in cats receiving empty liposome vehicle. CONCLUSIONS: SSL-CDDP appears to be safe to administer to cats at a dosage of 70 mg of free CDDP equivalent/m2, a CDDP dose known to be therapeutic in dogs. Pyrexia, although marked, appears to be a short-term and well tolerated side effect. CLINICAL RELEVANCE: SSL-CDDP appears to abrogate the uniformly fatal side effects associated with administration of tumoricidal quantities of free CDDP to cats. This new formulation should allow investigation of the antitumor properties of CDDP against spontaneously arising neoplasms in cats.

Animals↗

Evaluation of analgesia provided by postoperative administration of butorphanol to cats undergoing onychectomy.

OBJECTIVE: To evaluate adequacy of analgesia provided by postoperative administration of butorphanol to cats undergoing onychectomy. DESIGN: Randomized controlled trial. ANIMALS: 63 cats undergoing elective onychectomy. PROCEDURE: Cats were randomly assigned to a treatment (n = 42) or control group (21). Cats in the treatment group were given butorphanol parenterally immediately and 4 hours after surgery and orally for 2 days after surgery. Rectal temperature, heart rate, and respiratory rate were recorded and scores were assigned for temperament, recovery, sedation, analgesia, and lameness for the first 24 hours after surgery. Owners provided scores for appetite, personality, and lameness the first and second days after discharge from the hospital. RESULTS: Heart rate, respiratory rate, rectal temperature, and temperament and sedation scores were not significantly different between groups at any evaluation time. Recovery scores were significantly better for butorphanol-treated than for control-group cats 10 minutes after extubation. Analgesia scores were significantly better for butorphanol-treated than for control-group cats between 5 and 24 hours after surgery. Fewer butorphanol-treated than control-group cats were lame at the time of discharge from the hospital. The first day after discharge, owners reported that percentages of butorphanol-treated cats that ate normally, acted normally, and had only mild or no lameness were significantly higher than percentages of control-group cats that did. Significant differences between groups were not detected the second day after discharge. CLINICAL IMPLICATIONS: Results suggest that for cats undergoing onychectomy, administration of butorphanol the day of surgery and the first full day after surgery provides effective analgesia and improves recovery, appetite, and gait.

Analgesia↗

Evidence of reproductive failure and lack of perinatal transmission of Bartonella henselae in experimentally infected cats.

Five female specific pathogen-free (SPF) cats inoculated intradermally with B. henselae and bacteremic for 4 weeks, and one cat inoculated with 0.9% NaCl, were bred with uninfected SPF male cats. The uninfected female became pregnant with one breeding, while three infected cats became pregnant 1-12 weeks later, after repeated breedings. Two infected females either did not become pregnant or maintain pregnancies despite repeated breedings. Infected cats produced anti-B. henselae IgM and IgG antibodies. Fetuses and kittens of infected cats were not infected and did not produce anti-B. henselae antibodies. Male cats bred with infected females did not become infected or seroconvert. Maternal anti-B. henselae IgG antibodies detected in sera of kittens 2 weeks post-partum were no longer detectable 10 weeks post-partum. These findings suggest that B. henselae causes reproductive failure in female cats, but is not transmitted transplacentally, in colostrum or milk, or venereally. Infected cats immunosuppressed with methylprednisolone acetate after their kittens were weaned had no detectable bacteria in tissues, suggesting that they were no longer infected.

Animals↗

Low prevalence of Bartonella henselae infections in Norwegian domestic and feral cats.

Bartonella henselae is the causative agent of cat scratch disease (CSD). This clinical entity is very rarely encountered in human medical practice in Norway. B. henselae infections including bacteraemia in cats have been frequently reported. The objective of the present study was to investigate the seroprevalence rate and the degree of B. henselae bacteraemia in Norwegian domestic and feral cats. One hundred cats investigated at a small animal veterinary practice in the middle of Norway were included in the study. Blood collected in Isolator blood-lysis tubes and lysates of erythrocytes after freezing and thawing were cultured. PCR analysis of whole blood was also performed. Serology was performed by indirect fluorescence assay (IFA) and enzyme immunoassay (EIA) using immobilised B. henselae Houston-1 strain as antigen. None of the 100 cats investigated was found to be bacteraemic. All 100 cats were seronegative when analysed by IFA; one cat was positive by EIA. The discrepancy between IFA and EIA of this particular cat is probably due to cross-reactive antibodies. Contrary to findings reported from several geographic regions, B. henselae infections in Norwegian cats appear to be virtually absent. This in turn may explain why CSD has not been reported in human medical practice in Norway.

Animals↗

Coinfection with Bartonella clarridgeiae and Bartonella henselae and with different Bartonella henselae strains in domestic cats.

Bartonella clarridgeiae and several strains of Bartonella henselae, the agent of cat scratch disease, with variations in the 16S rRNA gene have been found to infect the blood of cats. An epidemiologic study of Bartonella infection in domestic French cats revealed that of 436 cats sampled, 5 cats (1.1%) were coinfected with B. henselae and B. clarridgeiae and 2 cats (0.5%) were coinfected with two strains of B. henselae with variations in the 16S rRNA gene, B. henselae type I and type II. In an indirect immunofluorescence assay, coinfected cats tested positive for both Bartonella species at titers of > or = 128. Identification of the colonies was achieved by preformed enzyme analysis, PCR-restriction fragment length polymorphism analysis of the citrate synthase gene, and 16S rRNA gene sequencing. Colony size differences in mixed culture allowed differentiation of the Bartonella species. The coinfection of cats with two Bartonella species or variants of the same species raises concern about the possibility of dual infection in humans. The development of a polyvalent vaccine targeted against the most pathogenic or invasive strains may be a means of protecting cats and man from infection.

Animals↗

Cellular expression of an HMGCR promoter-CAT fusion gene in transgenic mouse brain: evidence for a developmental regulation in oligodendrocytes.

The HMGCR gene encodes the 3-hydroxy-3-methylglutaryl coenzyme A reductase, which is the key enzyme for cholesterol synthesis. Mice transgenic for the prokaryotic chloramphenicol acetyl transferase (CAT) reporter gene fused with a 5' Bam H1 fragment including the promoter sequence for murine HMGCR gene have been obtained. Homozygote transgenic mice were derived from a particular line selected for similar regulation of endogenous HMGCR and the transgene expression by nutritional conditions in different tissue. In addition, high expression of the transgene was evidenced in the brain. Cellular expression of the CAT gene in the central nervous system (CNS) was investigated by immunohistochemistry (IHC). This study was performed on frozen sections of the developing and adult brain, using a rabbit anti-CAT antiserum especially raised for that purpose. CAT expression was observed in some rare individuals in different neural cell types including Purkinje cells and astrocytes. But the most outstanding observation was the high level of CAT expression correlated with differentiated pattern of oligodendrocyte (Ol) distribution observed in white-matter tracts. Double and triple labeling for CAT and stage-specific antigens were performed on transgenic Ol-enriched preparations and cultures. This study showed a normal sequence of differentiation in the transgenic oligodendroglial cell lineage and demonstrated a strict correlation between late differentiation and activation of the CAT gene in these cells: CAT expression started in transgenic Ols between galactocerebroside (GC)-positive and myelin basic protein (MBP)-positive stages and was detected in MBP-positive cells during the myelination period. After myelination, the number of CAT-positive Ols decreased in the adult brain. These observations demonstrate a developmental regulation of the CAT transgene in Ols during myelination in CNS and reinforce the hypothesis of endogenous synthesis as major source of cholesterol during myelination.

Animals↗

A transcription termination signal immediately precedes the coding sequence for the chloramphenicol-inducible plasmid gene cat-86.

The plasmid gene cat-86 specifies chloramphenicol-inducible, chloramphenicol acetyltransferase in Bacillus subtilis. The inducible regulation is independent of the promoter that is used to activate cat-86 and is independent of the cat-86 coding sequence. We have proposed that the regulation of cat-86 results from the transcription of a pair of inverted-repeat sequences that immediately precede the coding sequence. These transcripts are predicted to sequester the cat-86 ribosome binding site in a stable RNA stem-loop which, in theory, should block the ribosome binding site from pairing with 16S rRNA. Inducible expression of cat-86 may therefore result in part from regulation of the translation of cat-86 mRNA. However, chloramphenicol-induction correlates with increased levels of cat-86 mRNA and the RNA stem-loop preceding the cat-86 coding sequence structurally resembles a rho-independent transcription terminator. We have therefore tested the inverted-repeats as a potential site of transcription termination. Transcription studies performed in vitro using SP6 RNA polymerase and in vivo by S1 mapping demonstrate that a substantial fraction of the potential cat-86 transcripts terminate at a site immediately 3' to the inverted-repeats. The results of the in vivo experiments suggest that the termination signal may be partially relieved by growth of cells in chloramphenicol.

Acetyltransferases↗

Concurrence of cat and tet genes in multiple antibiotic-resistant bacteria isolated from a sea cucumber and sea urchin mariculture farm in China.

A basic understanding of abundance and diversity of antibiotic-resistant microbes and their genetic determinants is necessary for finding a way to prevent and control the spread of antibiotic resistance. For this purpose, chloramphenicol and multiple antibiotic-resistant bacteria were screened from a mariculture farm in northern China. Both sea cucumber and sea urchin rearing ponds were populated with abundant antibiotic-resistant bacteria, especially marine vibrios. Sixty-five percent chloramphenicol-resistant isolates from sea cucumber harbored a cat gene, either cat IV or cat II, whereas 35% sea urchin isolates harbored a cat gene, actually cat II. The predominant resistance determinant cat IV gene mainly occurred in isolates related to Vibrio tasmaniensis or Pseudoalteromonas atlantica, and the cat II gene mainly occurred in Vibrio splendidus-like isolates. All the cat-positive isolates also harbored one or two of the tet genes, tet(D), tet(B), or tet(A). As no chloramphenicol-related antibiotic was ever used, coselection of the cat genes by other antibiotics, especially oxytetracycline, might be the cause of the high incidence of cat genes in the mariculture farm studied.

Animals↗

Contraceptive potential of porcine zona pellucida in cats.

The contraceptive potential of solubilized porcine zona pellucida (spZP) was studied in 2 groups of cats after active immunization using slightly different protocols. Cats from Group 1 (n = 3) were immunized with a total of 300 8g spZP divided in 4 s.c. multisite injections (each of 37.5 8g) given at 10 day intervals followed by a booster 150 days after the initial immunization. Cats from Group 2 (n = 5) were immunized with a total of 400 8g spZP divided in 4 i.m. injections (each of 50 8g) given at 2 wk intervals followed by a booster 92 days after initial immunization. Immunogen was emulsified in Complete Freund Adjuvant for the first dose and in Incomplete Freund Adjuvant for the following 3 doses. The respective controls were immunized in the same manner using only adjuvant and PBS. Immunofluorescence studies showed intense fluorescence on the zona pellucida (ZP) of the oocytes isolated from immunized cats, as well as on the ZP of porcine and cat oocytes preincubated in immune sera. Sera from cats immunized with spZP inhibited in vitro binding was demonstrated in oocytes isolated from immunized group 1 cats. In vivo fertility data in Group 2 cats revealed that only 1 of 5 cats became pregnant, the one with the lowest anti-spZP titer. The results from the experiments reported above, suggest that in this preliminary study spZP can elicit antibodies with contraceptive potential in actively immunized female cats.

Journal Article↗

Chloramphenicol induction of cat-86 requires ribosome stalling at a specific site in the leader.

The plasmid gene cat-86 specifies chloramphenicol-inducible chloramphenicol acetyltransferase in Bacillus subtilis. Induction by the antibiotic is primarily due to activation of the translation of cat-86-encoded mRNA. It has been suggested that the inducer stalls ribosomes at a discrete location in the leader region of cat-86 mRNA, which causes the destabilization of a downstream RNA secondary structure that normally sequesters the cat-86 ribosome binding site. It is the destabilization of this RNA secondary structure that permits translation of the cat-86 coding sequence. In the present report, we show that ribosomes that were stalled in the cat-86 leader by starvation of host cells for the amino acid specified by leader codon 6 induced gene expression to a level above that detected when cells were starved for the amino acids specified by leader codons 7 and 8. Starvation for amino acids specified by leader codons 3, 4, or 5 failed to activate cat-86 expression. These results indicate that the stalled ribosome that is most active in cat-86 induction has its aminoacyl site occupied by leader codon 6. To determine if chloramphenicol also stalled ribosomes in the cat-86 regulatory leader such that the aminoacyl site was occupied by codon 6, we separately changed leader codons 3, 4, 5, and 6 to the translation termination (ochre) codon TAA. Each of the mutated genes was tested for its ability to be induced by chloramphenicol. The results show that replacement of leader codons 3, 4, or 5 by the ochre codon blocked induction, whereas replacement of leader codon 6 by the ochre codon permitted induction. Collectively, these observations lead to the conclusion that cat-86 induction requires ribosome stalling in leader mRNA, and they identify leader codon 6 as the codon most likely to be occupied by the aminoacyl site of a stalled ribosome that is active in the induction.

Acetyltransferases↗

Perturbing highly conserved spatial relationships in the regulatory domain that controls inducible cat translation.

Chloramphenicol activates translation of cat-86 mRNA by stalling a ribosome in the leader of individual transcripts. Stalling triggers two sequential events: the destabilization of a region of secondary structure that sequesters the cat ribosome-binding site (RBS-C), and the initiation of cat translation. The site of drug-dependent ribosome stalling is dictated by the leader sequence, crb; crb causes a ribosome to stall with its aminoacyl site at leader codon 6. We demonstrate that induction requires the maintenance of a precise spatial relationship between crb and sequences within the left inverted repeat of the secondary structure. Therefore, destabilization of the secondary structure during chloramphenicol induction may result from the interaction of a stalled ribosome with a specific sequence in the secondary structure rather than from non-specific masking of RNA sequences. cat-86 regulation also depends on the distance that separates crb from RBS-C. This interval of 33 nucleotides was incrementally increased and decreased by mutations within a loop in the secondary structure. Shortening the distance between crb and RBS-C by three nucleotides reduced induction by half and a deletion of nine nucleotides abolished induction. Insertion mutations were without effect on induced expression but elevated basal expression. The results indicate that when the A site of a ribosome occupies leader codon 6 the secondary structure is destabilized and there is no interference with entry of a second ribosome at RBS-C. The data further demonstrate that when the A site of a ribosome in the leader is within 30 nucleotides of RBS-C, cat expression decreases. This decrease probably results from competition of the leader ribosome with the ribosome initiating cat translation. Our observations demonstrate that in wild-type cat-86 the distances between crb and the secondary structure, and between crb and RBS-C provide the precise spacing necessary to achieve three interdependent effects: the destabilization of the RNA secondary structure by a ribosome stalled at crb; a lack of competition between a ribosome stalled at crb and the initiating ribosome; and maintenance of a low, but measurable, basal level of cat expression. The spatial relationships identified as necessary for the regulation of cat-86 are conserved in the regulatory regions for five other inducible cat genes.

Base Sequence↗