Klebsiella pneumoniae Structural Genomics

Collaboration with Prof. Huang, Tai-Huang,
High-field Macromolecular NMR Core Facility of the National Research Program for Genomic Medicine.

Klebsiella pneumoniae is a gram-negative enteric rod and a well-known pathogen of nosocomial infections and infections in compromised hosts. In the past 15 years, however, a new type of invasive K. pneumoniae disease has emerged in Taiwan that typically presents as community-acquired primary liver abscess in patients without biliary tract diseases or other intra-abdominal infections. K. pneumoniae is the only pathogen can be identified. Sepsis syndrome and bacteremia are common, with metastatic meningitis or septic endophthalmitis complicates the course in 10-12% of cases. Diabetes mellitus, which can be a predisposing factor, can be detected in about 50% of the patients, but the rest had no apparent underlying diseases. Another presentation of this invasive K. pneumoniae disease is community-acquired meningitis. There was a gradual increase in community-acquired meningitis due to K. pneumoniae in Taiwan since 1987. After 1996, K. pneumoniae replaced Streptococcus pneumoniae as the leading pathogen of community-acquired bacterial meningitis in adult. Although the responsible K. pneumoniae strains were uniformly susceptible to all cephalosporins and aminoglycosides, mortality as high as 10-20% for primary liver abscess and as high as 30íV40% among those with meningitis has been reported. Invasive K. pneumoniae disease in Taiwan is a new threat to public health and deserves further attention and studies.

Structural genomics is a new and rapidly developing field in life sciences. The goal of this field is to discover and analyze the structures as well as functions of all proteins in nature in order to provide a foundation for a fundamental understanding of biology. Several genome projects have been finished recently. The next step of genomic studies after the yielding of the complete genome sequence of a species is to identify both cellular and molecular function of each gene in the genome. Several pilot projects in structural genomics are on the way. The preliminary results from these pilot projects show complementary data with functional genomic analysis. Both approaches are important for genome research beyond DNA sequencing. In this program project, we propose to develop and incorporate the methodologies and technologies for structural genomics and proteomics studies on Klebsiella pneumoniae and its associated proteins involved in cell cycle.

This bioinformatics web site is for Klebsiella pneumoniae structural genomics (http://kp.life.nthu.edu.tw/). With the help from bioinformatics, high throughput cloning, expression, and purification of target proteins, structural analysis by various biophysical techniques and proteomics studies on the cellular function of Klebsiella pneumoniae, we anticipate to solve about 10-20 novel protein structures each year. In addition to the structural genomics studies, specific proteins for cellular responses induced by Klebsiella pneumoniae or its major virulence factors by comparative studying of differential protein expression profiling both in sub-cellular organelles and whole cells will be studied by using the proteomics approaches such as 2D gel, mass spectroscopy, confocal microscopy, etc. The completion of this project will expand our understanding of the atomic basis of Klebsiella pneumoniae, provide significant shortcuts to understanding its gene function, and generate the most fundamental genomic data besides the sequence information itself. The structure-function information would ultimately aid in understanding the molecular mechanism of cellular response to Klebsiella pneumoniae infection. Furthermore, the findings from this project may provide insight to the prevention and treatment of diseases associated with Klebsiella pneumoniae.