Serino G, Maliga P
Waksman Institute, Rutgers, The State University of New Jersey, 190 Frulinghuysen Road, Piscataway, New Jersey 08854-8020, USA.
Plant Physiol. 1998 Aug;117(4):1165-70. doi: 10.1104/pp.117.4.1165.
Plastid genes in photosynthetic higher plants are transcribed by at least two RNA polymerases. The plastid rpoA, rpoB, rpoC1, and rpoC2 genes encode subunits of the plastid-encoded plastid RNA polymerase (PEP), an Escherichia coli-like core enzyme. The second enzyme is referred to as the nucleus-encoded plastid RNA polymerase (NEP), since its subunits are assumed to be encoded in the nucleus. Promoters for NEP have been previously characterized in tobacco plants lacking PEP due to targeted deletion of rpoB (encoding the beta-subunit) from the plastid genome. To determine if NEP and PEP share any essential subunits, the rpoA, rpoC1, and rpoC2 genes encoding the PEP alpha-, beta'-, and beta"-subunits were removed by targeted gene deletion from the plastid genome. We report here that deletion of each of these genes yielded photosynthetically defective plants that lack PEP activity while maintaining transcription specificity from NEP promoters. Therefore, rpoA, rpoB, rpoC1, and rpoC2 encode PEP subunits that are not essential components of the NEP transcription machinery. Furthermore, our data indicate that no functional copy of rpoA, rpoB, rpoC1, or rpoC2 that could complement the deleted plastid rpo genes exists outside the plastids.
光合高等植物中的质体基因由至少两种RNA聚合酶转录。质体rpoA、rpoB、rpoC1和rpoC2基因编码质体编码的质体RNA聚合酶(PEP)的亚基,这是一种类似于大肠杆菌的核心酶。第二种酶被称为核编码的质体RNA聚合酶(NEP),因为其亚基被认为是由核编码的。由于从质体基因组中靶向缺失rpoB(编码β亚基),NEP的启动子先前已在缺乏PEP的烟草植物中得到表征。为了确定NEP和PEP是否共享任何必需亚基,通过从质体基因组中靶向基因缺失,去除了编码PEPα-、β'-和β"-亚基的rpoA、rpoC1和rpoC2基因。我们在此报告,这些基因中的每一个的缺失都产生了光合缺陷植物,这些植物缺乏PEP活性,同时保持了来自NEP启动子的转录特异性。因此,rpoA、rpoB、rpoC1和rpoC2编码的PEP亚基不是NEP转录机制的必需组成部分。此外,我们的数据表明,质体外不存在能够补充缺失的质体rpo基因的rpoA、rpoB、rpoC1或rpoC2的功能拷贝。