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The genomic DNA of bacteria is highly compacted in a single or a few bodies known as nucleoids. Here, we have isolated Escherichia coli nucleoid by sucrose density gradient centrifugation. The sedimentation rates, structures as well as protein/DNA composition of isolated nucleoids were then compared under various growth phases. The nucleoid structures were found to undergo changes during the cell growth; i. e., the nucleoid structure in the stationary phase was more tightly compacted than that in the exponential phase. In addition to factor for inversion stimulation(Fis), histone-like nucleoid structuring protein(H-NS), heat-unstable nucleoid protein(HU) and integration host factor(IHF) here we have identified, three new candidates of E. coli nucleoid, namely DNA-binding protein from starved cells(Dps), host factor for phage Qβ(Hfq) and suppressor of td- phenotype A(Stp A). Our results reveal that the major components of exponential phase nucleoid are Fis, HU, H-NS, Stp A and Hfq, while Dps occupies more than half of the stationary phase nucleoid. It has been known for a while that Dps is the main nucleoid-associated protein at stationary phase. From these results and the prevailing information, we propose a model for growth phase dependent changes in the structure and protein composition of nucleoid in E. coli.
The genomic DNA of bacteria is highly compacted in a single or a few bodies known as nucleoids. Here, we have isolated Escherichia coli nucleoid by sucrose density gradient centrifugation. The sedimentation rates, structures as well as protein / DNA composition of isolated nucleoids were then compared under various growth phases. The nucleoid structures were found to undergo changes during the cell growth; ie, the nucleoid structure in the stationary phase was more tightly compacted than that in the exponential phase. histone-like nucleoid structuring protein (H-NS), heat-unstable nucleoid protein (HU) and integration host factor (IHF) here we have identified, three new candidates for E. coli nucleoid, ie DNA- binding protein from starved cells Dps), host factor for phage Qβ (Hfq) and suppressor of td-phenotype A (Stp A). Our results reveal that the major components of exponential phase nucleoids are Fis, HU, H-NS, Stp A and Hfq, whi le Dps occupies more than half of the stationary phase nucleoid. It has been known for a while that Dps is the main nucleoid-associated protein at stationary phase. From these results and the prevailing information, we propose a model for growth phase dependent changes in the structure and protein composition of nucleoid in E. coli.