Sequences in the -35 region of Escherichia coli rpoS-dependent genes promote transcription by E sigma S.

sigma S is an alternate sigma factor which functions with RNA polymerase to activate transcription of genes that are involved in a number of stress responses, including stationary-phase survival and osmoprotection. The similarity of the sigma S protein to sigma D (Escherichia coli's major sigma factor) in the regions thought to recognize and bind promoter sequences suggests that sigma S- and sigma D-associated RNA polymerases recognize promoter DNA in a similar manner. However, no promoter recognition sequence for sigma S holoenzyme (E sigma S) has been identified. An apparent conservation of cytosine nucleotides was noted in the -35 region of several sigma S-dependent promoters. Site-directed mutagenesis and reporter gene fusions were used to investigate the importance of the -35 cytosine nucleotides for sigma S-dependent transcription. Substitution of cytosine nucleotides for thymidine at the -35 site of the sigma D-dependent proU promoter effectively abolished transcription by E sigma D but allowed E sigma S to direct transcription from the mutant promoter. Inclusion of the sigma D consensus -10 hexamer strengthened transcription by E sigma S, demonstrating that both E sigma D and E sigma S can recognize the same -10 sequences. Conversely, replacement of -35 site cytosine nucleotides with thymidine in the sigma S-dependent osmY promoter reduced transcription by E sigma S and increased transcription by E sigma D. Our data suggest that DNA sequences in the -35 region function as part of a discriminator mechanism to shift transcription between E sigma D and E sigma S.

The Escherichia coli proU promoter element and its contribution to osmotically signaled transcription activation.

The proU operon of Escherichia coli encodes a high-affinity glycine betaine transport system which is osmotically inducible and enables the organism to recover from the deleterious effects of hyperosmotic shock. Regulation occurs at the transcriptional level. KMnO4 footprinting showed that the preponderance of transcription initiated at a single primary promoter region and that proU transcription activation did not occur differentially at alternate promoters in response to various levels of salt shock. Mutational analysis confirmed the location of the primary promoter and identified an extended -10 region required for promoter activity. Specific nucleotides within the spacer, between position -10 and position -35, were important for maximal expression, but every mutant which retained transcriptional activity remained responsive to osmotic signals. A chromosomal 90-bp minimal promoter fragment fused to lacZ was not significantly osmotically inducible. However, transcription from this fragment was resistant to inhibition by salt shock. A mutation in osmZ, which encodes the DNA-binding protein H-NS, derepressed wild-type proU expression by sevenfold but did not alter expression from the minimal promoter. The current data support a model in which the role of the proU promoter is to function efficiently at high ionic strength while other cis-acting elements receive and respond to the osmotic signal.

Molecular characterization of the promoter of osmY, an rpoS-dependent gene.

The osmY gene, which encodes a periplasmic protein with an apparent M(r) of 22,000, is induced by both osmotic and growth phase signals. We demonstrate here that osmY expression is regulated at the level of transcription and that transcription initiates 242 nucleotides upstream of the osmY open reading frame. Relative to the transcriptional start site, 5' deletions up to -36 did not inhibit osmY expression. 3' deletions that extended into the untranslated leader region affected the overall level of osmY::lacZ expression but did not affect inducibility. 5' and 3' deletions that extended past the transcriptional start region essentially abolished osmY expression, suggesting that there is a single promoter region. A putative promoter was identified, and its -10 region, TATATT, closely resembles the sigma 70 consensus -10 sequence, TATAAT. However, we show that osmY is not absolutely dependent on a functional sigma 70 for its expression. Since osmY expression does require rpoS (R. Hengge-Aronis, R. Lange, N. Henneberg, and D. Fischer, J. Bacteriol. 175:259-265, 1993), which encodes a stationary-phase sigma factor, sigma S (K. Tanaka, Y. Takayanagi, N. Fujita, A. Ishihama, and H. Takahashi, Proc. Natl. Acad. Sci. USA 90:3511-3515, 1993), E sigma S may be the form of RNA polymerase responsible for transcription of osmY.

Decentration of flexible loop posterior chamber intraocular lenses in a series of 222 postmortem eyes.

Two hundred twenty-two postmortem eyes containing posterior chamber intraocular lenses (IOLs) were analyzed for optic decentration in relationship to lens style, implant duration, and loop fixation site. Decentration values were not affected significantly by either lens style or implant duration. In 33.3% of specimens, both loops were situated within the lens capsular sac, 18.0% had both loops fixated in the ciliary sulcus, and in 48.7% one loop was fixated in the lens capsular sac and the opposite loop in the ciliary sulcus or zonular region. There was a statistically significant difference in the amount of decentration in the three fixation groups studied. Capsular fixation provides the best and most consistent centration compared with fixation of both loops in the ciliary sulcus or asymmetrical fixation with only one loop in the capsular sac.

Visual aberrations caused by optic components of posterior chamber intraocular lenses.

A 58-year-old airline pilot had cataract surgery with implantation of a posterior chamber intraocular lens with four positioning holes around the optic edge. During periods of maximum pupil dilation, such as at night, visual aberrations including glare, monocular diplopia, and haloes occurred and he was unable to work in his occupation. The symptoms were severe enough that lens exchange was required, and a posterior chamber lens with no positioning holes was successfully implanted. The symptoms immediately subsided postoperatively and his last known visual acuity was 20/15. This case, and the report of another patient with similar postoperative problems, illustrates that implantation of lens optics with a larger effective optical zone for posterior chamber lens implantation is desirable. This is particularly true now that younger, more active patients, many still engaged in occupations, are having lens implantations.

Anterior chamber lenses. Part I: Complications and pathology and a review of designs.

Clinicopathologic data to assist ophthalmologists in choosing a safe and efficacious anterior chamber intraocular lens (IOL) are rapidly becoming available. Two important factors that have led to an increased success rate with some anterior chamber IOL styles are (1) attention to lens design and (2) attention to modern manufacturing and lens finishing techniques. We now know much more about how to achieve appropriate lens flexibility, which decreases the need for perfect sizing. Increased attention has been given to the anterior-posterior vaulting characteristics of IOLs. This has reduced the incidence of various complications such as the intermittent touch syndrome and the uveal chafing syndrome. We recognize several design flaws in some lens styles. For example, there is now a considerable decrease in the number of small-diameter, round-looped anterior chamber IOLs being implanted, particularly those with a closed-loop configuration. Several problems have been and continue to be caused by some poorly manufactured anterior chamber lenses with sharp optic and haptic edges. Technology to assure smooth lens finishing and polishing is available and readily accessible to all manufacturers. Defective lenses should soon be a thing of the past.

Posterior chamber intraocular lenses in a series of 75 autopsy eyes. Part II: Postimplantation loop configuration.

We illustrate, from the pathologist's viewpoint, the configuration of posterior chamber lens loops within the eye. The final configuration of these loops is often a "C" shape, particularly following capsular fixation. When one or both loops are placed in the ciliary sulcus, where tissue resistance to loop compressive forces is less than in the capsular sac, an invagination or erosion of the loop into the ciliary body stroma or ciliary muscle frequently occurs. Although a surgeon should first and foremost choose a lens design with which he or she is comfortable in terms of ease of implantation and good clinical results, our pathological observations indicate that a subtle compromise in loop configuration might provide a more physiologically correct configuration in the eye, particularly for those who prefer in-the-bag implantation. Most surgeons prefer a lens design that will provide adequate fixation in the ciliary sulcus if an attempted capsular sac implantation is not successful.

Posterior chamber intraocular lenses in a series of 75 autopsy eyes. Part III: Correlation of positioning holes and optic edges with the pupillary aperture and visual axis.

The possibility that undesirable visual complications such as glare, halo, monocular diplopia, or other visual aberrations can result from the presence of posterior chamber lens optic edges or such lens elements as positioning holes or loop-optic junctions within the pupillary aperture has received little attention. There is recent clinical evidence that these phenomena may be clinically significant. In a series of 75 autopsy eyes with posterior chamber intraocular lenses, we have observed that in 71% of cases an optic edge, or element of the optic such as a positioning hole, was situated either within the pupillary aperture and visual axis (average pupillary diameter 3.45 mm) or within 0.5 mm of the pupillary margin. This finding was most common (92%) in cases with asymmetric placement, less common (50%) in cases with symmetric placement. Many more young patients are now undergoing implantation surgery. These patients generally have wider, more mobile pupils, and they may be more aware of subjective symptoms, particularly at night. Subtle changes in implantation techniques and in lens design and manufacture can minimize complications related to this condition. These changes include symmetric loop placement (both loops in the capsular sac or both in the ciliary sulcus) to decrease optic decentration. Other measures such as making partial-depth positioning holes, increasing the size of the optic, eliminating or reducing the number of positioning holes, and placing positioning holes in tabs on the optic edges may function to increase the effective optical zone.

Posterior chamber intraocular lenses in a series of 75 autopsy eyes. Part I: Loop location.

Over a period of 27 months, November 1983 to February 1986, 75 eyes obtained postmortem with posterior chamber intraocular lenses (IOLs) were examined at the Center for Intraocular Lens Research, University of Utah Health Sciences Center. These IOLs were studied by histopathological techniques to determine the location of the loops. The most common combination, found in 47% of the specimens, was one loop in the lens capsular sac (bag) and one loop in the ciliary sulcus. In 32% of the specimens, both loops were in the capsular sac; in 17%, both loops were in the ciliary sulcus. Compared to results observed in other autopsy studies, in which capsular fixation was documented in less than 3% of cases, these findings reflect a trend toward capsular sac (in-the-bag) implantation of open-looped posterior chamber IOLs.