Cell cycle expression of the heat shock groESL operon in Caulobacter crescentus

Author(s)

Regina Lúcia Baldini

• Document type: Doctoral Thesis
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Conjunto das Químicas (IQ e FCF) (CQ/DBDCQ)
• Defense date: 1999-04-23

Examining board members

Suely Lopes Gomes; Gabriel Padilla Maldonado; Carlos Frederico Martins Menck; Carla Columbano de Oliveira; Roberto Vicente Santelli

• Advisor: Suely Lopes Gomes

Abstract

Caulobacter crescentus is an aquatic free-living Gram-negative bacterium whose cell cycle depends on cell differentiation. The asymmetrical predivisional cell gives rise to two morphologically and functionally different daughter cells: the swarmer cell an the stalked cell. The expression of the groESL operon is induced by heat shock and is cell cycle controlled at normal temperatures, with maximal transcription in the predivisional cell and very low levels in the stalked cell. In this work, it was demonstrated that, in a strain overexpressing σ32, the levels of groESL transcripts and the synthesis of GroEL are increased, confirming that this factor is responsible for the transcriptional activation of the σ32 -like promoter of this operon, that also presents a inverted repeat called CIRCE in its regulatory region. groESL-lacZ transcription fusions with point mutations in CIRCE indicated a negative role of this cis-acting element only at normal growth temperatures, with a minor effect on heat shock induction. In addition, the expression of these fusions are no longer cell-cycle regulated, as well as GroEL synthesis in a strain which does not have the HrcA protein, the putative repressor that binds CIRCE, indicating that the CIRCE-HrcA system are involved in cell cycle regulation of groESL in C. crescentus. It was also shown that groEL is an essential gene at normal growth temperatures, since a strain with groEL disrupted is not viable. A conditional mutant was obtained by homologous recombination and in restrictive conditions growth is inhibited, DnaK levels are increased and the cells become filamentous, but no celllysis was observed. The proteins that require GroEL/GroES for proper folding have not been identified yet. (AU)