Small subunits of RNA polymerase: localization, levels and implications for core enzyme composition Free

Abstract

Bacterial RNA polymerases (RNAPs) contain several small auxiliary subunits known to co-purify with the core , and ′ subunits. The subunit is conserved between Gram-positive and Gram-negative bacteria, while the subunit is conserved within, but restricted to, Gram-positive bacteria. Although various functions have been assigned to these subunits via assays, very little is known about their roles. In this work we constructed a pair of vectors to investigate the subcellular localization of the and subunits in with respect to the core RNAP. We found these subunits to be closely associated with RNAP involved in transcribing both mRNA and rRNA operons. Quantification of these subunits revealed to be present at equimolar levels with RNAP and to be present at around half the level of core RNAP. For comparison, the localization and quantification of RNAP ′ and subunits in was also investigated. Similar to , ′ and closely associated with the nucleoid and formed subnucleoid regions of high green fluorescent protein intensity, but, unlike in , levels in were close to parity with those of ′. These results indicate that is likely to be an integral RNAP subunit in Gram-positives, whereas levels differ substantially between Gram-positives and -negatives. The subunit may be required for RNAP assembly and subsequently be turned over at different rates or it may play roles in Gram-negative bacteria that are performed by other factors in Gram-positives.

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2010-12-01
2024-03-28
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