Biomedical subjects

J J Kohler

Publications and source records attributed to J J Kohler.

At least 19 recordsLinked to original sources

PubMed2001

Kinetic studies of Fos.Jun.DNA complex formation: DNA binding prior to dimerization.

The bZIP proteins Fos and Jun bind DNA rapidly and with high affinity, forming a heteromeric complex that mediates activated transcription. Here we use stopped-flow fluorescence resonance energy transfer (FRET) to study the kinetic pathway by which Fos.Jun. DNA complexes assemble. Though dimerization of Fos and Jun occurs rapidly in the absence of DNA, the rate of dimerization is enhanced in the presence of DNA. Global analysis of the kinetic data shows that the favored DNA binding pathway is one is which the two protein monomers bind DNA sequentially and assemble their dimerization interface while bound to DNA.

Amino Acid Sequence↗

PubMed2001

Effects of nucleic acids and polyanions on dimer formation and DNA binding by bZIP and bHLHZip transcription factors.

A large fraction of known transcription factors form 2:1 complexes with DNA. In our studies of the assembly of such ternary (protein-protein-DNA) complexes formed by bZIP and bHLHZip proteins, we found that the proteins recognize DNA as monomers. Here we show that protein monomer-DNA complexes are favored at high DNA concentrations. Further, we show that, due to fast rates of association with protein monomers, DNA and other polyanions accelerate the rate of protein dimer formation. Finally, we find that DNA-assisted formation of protein dimers provides a mechanism by which dimeric transcription factors can rapidly discriminate between specific and nonspecific sites.

Basic Helix-Loop-Helix Proteins↗

PubMed2000

Effect of preexisting immunity to Salmonella on the immune response to recombinant Salmonella enterica serovar typhimurium expressing a Porphyromonas gingivalis hemagglutinin.

Recombinant Salmonella strains expressing foreign heterologous genes have been extensively studied as live oral vaccine delivery vectors. We have investigated the mucosal and systemic immune responses following oral immunization with a recombinant Salmonella enterica serovar Typhimurium expressing the hemagglutinin HagB from Porphyromonas gingivalis, a suspected etiological agent of adult periodontal disease. We have previously shown a primary mucosal and systemic response following oral immunization with chi4072/pDMD1 and recall responses following boosting at 14 weeks after primary immunization. In this study, we examined the effects of earlier boosting as well as the effects of deliberately induced immunity to the Salmonella carrier strain on subsequent immune responses. Mice boosted at week 7 following immunization, a point which corresponded to the peak of the primary response, generally showed lower responses than those boosted at week 14. When mice were preimmunized with the Salmonella carrier alone and then immunized with the recombinant strain 7 or 14 weeks later, significant reductions were seen for serum immunoglobulin G (IgG) antibodies at week 14 and for salivary IgA at week 7. No reductions were seen in serum IgA or vaginal wash IgA antibodies. Mice appear to be refractory to boosting with orally administered salmonellae at 7 weeks. Deliberate immunization with the carrier strain did not appreciably affect recall responses at 14 weeks, with the exception of the serum IgG responses, nor did it affect colonization of the Peyer's patches.

Adhesins, Bacterial↗

PubMed2000

Long-term immunological memory induced by recombinant oral Salmonella vaccine vectors.

We have previously shown that Salmonella enterica serovar Typhimurium expressing the hagB hemagglutinin gene from Porphyromonas gingivalis can induce primary and recall immune responses in serum and secretions in mice; however, the longevity of memory induced by oral Salmonella carriers has not been adequately demonstrated. In this study, we examined the capacity of mice to mount a recall response 52 weeks after primary immunization. Recall responses were seen in serum immunoglobulin G (IgG) and IgA following boosting at week 52, and in most cases, they were equal to or greater than the primary responses. Significant mucosal IgA recall responses in saliva and vaginal wash were also detected following boosting at week 52. In addition, there was a considerable residual response in secretions at week 51, prior to boosting. These results indicate that oral Salmonella vectors can induce long-term memory to recombinant HagB and are particularly effective at inducing long-lasting mucosal responses as well as at inducing the capacity for mucosal recall responses.

Adhesins, Bacterial↗

PubMed1999

DNA specificity enhanced by sequential binding of protein monomers.

Transcriptional activation often requires the rapid assembly of complexes between dimeric transcription factors and specific DNA sites. Here we show that members of the basic region leucine zipper and basic region helix-loop-helix zipper transcription factor families follow an assembly pathway in which two protein monomers bind DNA sequentially and form their dimerization interface while bound to DNA. Nonspecific protein or DNA competitors have little effect on the rate of assembly along this pathway, but slow a competing pathway in which preformed dimers bind DNA. The sequential monomer-binding pathway allows the protein to search for and locate a specific DNA site more quickly, resulting in greater specificity prior to equilibrium.

Animals↗

PubMed1998

Oral immunization with recombinant Salmonella typhimurium expressing a cloned Porphyromonas gingivalis hemagglutinin: effect of boosting on mucosal, systemic and immunoglobulin G subclass response.

Live avirulent Salmonella typhimurium are convenient vaccine vectors for the delivery of recombinant antigens for the induction of mucosal and systemic immunity. The hagB gene encodes a hemagglutinin of Porphyromonas gingivalis, a suspected causal agent in human adult periodontal disease. In previous studies, we have shown that hagB can be expressed in avirulent S. typhimurium and is immunogenic when given orally to mice. In this study, we evaluated recall responses in both serum and mucosal secretions after boosting. In addition, we have examined the immunoglobulin G (IgG) subclass response in serum to both HagB and the Salmonella carrier. Mice were orally immunized with S. typhimurium expressing the hagB gene and then boosted 14 weeks later. Responses were measured through 27 weeks. Both primary and recall IgG and IgA responses were seen in serum to the purified HagB as well as to the Salmonella carrier. Likewise, mucosal primary and recall responses were seen in saliva, fecal extracts and vaginal washes although the kinetics of the responses differed. The anti-HagB response in serum was dominated by IgG2a during the peak of primary response, prior to boosting and during the peak of the recall response. The anti-S. typhimurium response shifted from predominantly IgG3 following primary immunization to IgG2a after boosting. The IgG1 response was minimal against each antigen. This pattern of IgG subclass distribution is consistent with a Th1-type response. These data indicate that avirulent S. typhimurium is capable of delivering a putative virulence factor from P. gingivalis and inducing a primary and recall response in both serum and secretions and provides a means of studying P. gingivalis virulence factors and for the development of a potential vaccine.

Administration, Oral↗

PubMed1975

[Villous tumors of the rectum. Apropos of 30 cases].

Although high localisations of villous tumours may be treated by wide removal, which alone guarantees cure without risk of relapse, the use of such an operation in low villous tumours is less obvious owing to the mutilation which may result. Local removal of small volume tumours by the anal route, by the trans-sacral route when they are larger or more broadly implanted, is preferred by us in rectal localisations, i.e. each villous tumour should be treated taking into considération all its characteristics.

Adult↗

PubMed1974