Malik M, Bensaid A, Rouviere-Yaniv J, Drlica K
Public Health Research Institute, New York, NY 10016, USA.
J Mol Biol. 1996 Feb 16;256(1):66-76. doi: 10.1006/jmbi.1996.0068.
The abundant bacterial protein called HU has the ability to wrap and bend DNA in vitro, and thus it has long been thought to play a role in DNA supercoiling. In the absence of HU, Escherichia coli formed tiny colonies on agar, rapidly accumulated suppressor mutations, and was hypersensitive to novobiocin. Three types of evidence implicated gyrase in the suppression of an HU deficiency. First, spontaneous suppressors that restored normal growth and reduced sensitivity to novobiocin mapped in gyrB, one of the genes encoding DNA gyrase. Second, a pair of known gyrB mutations (gyrB-203 Ts gyrB-221 NovR) allowed normal growth at permissive (30 degrees C) but not at intermediate (37 degrees C) conditions. Third, introduction of a gyrB-expressing plasmid restored normal colony size. DNA supercoiling comparisons showed that chromosomal supercoiling decreased in the absence of HU and increased toward wild-type levels in the presence of a spontaneous gyrB suppressor. Taken together, these data establish that HU has a physiological role in chromosomal DNA topology, probably by facilitating the action of gyrase.
一种名为HU的丰富细菌蛋白在体外具有包裹和弯曲DNA的能力,因此长期以来人们一直认为它在DNA超螺旋中发挥作用。在没有HU的情况下,大肠杆菌在琼脂上形成微小菌落,迅速积累抑制突变,并且对新生霉素高度敏感。有三类证据表明回旋酶参与了对HU缺陷的抑制。第一,恢复正常生长并降低对新生霉素敏感性的自发抑制子定位于gyrB,gyrB是编码DNA回旋酶的基因之一。第二,一对已知的gyrB突变(gyrB-203 Ts gyrB-221 NovR)在允许温度(30摄氏度)下能实现正常生长,但在中间温度(37摄氏度)条件下则不能。第三,引入一个表达gyrB的质粒可恢复正常菌落大小。DNA超螺旋比较表明,在没有HU的情况下染色体超螺旋减少,而在存在自发gyrB抑制子时则朝着野生型水平增加。综上所述,这些数据表明HU在染色体DNA拓扑结构中具有生理作用,可能是通过促进回旋酶的作用来实现的。