Activation of mRNA translation by phage protein and low temperature: the case of Lactococcus lactis abortive infection system AbiD1.
BACKGROUND
Abortive infection (Abi) mechanisms comprise numerous strategies developed by bacteria to avoid being killed by bacteriophage (phage). Escherichia coli Abis are considered as mediators of programmed cell death, which is induced by infecting phage. Abis were also proposed to be stress response elements, but no environmental activation signals have yet been identified. Abis are widespread in Lactococcus lactis, but regulation of their expression remains an open question. We previously showed that development of AbiD1 abortive infection against phage bIL66 depends on orf1, which is expressed in mid-infection. However, molecular basis for this activation remains unclear.


RESULTS
In non-infected AbiD1+ cells, specific abiD1 mRNA is unstable and present in low amounts. It does not increase during abortive infection of sensitive phage. Protein synthesis directed by the abiD1 translation initiation region is also inefficient. The presence of the phage orf1 gene, but not its mutant AbiD1R allele, strongly increases abiD1 translation efficiency. Interestingly, cell growth at low temperature also activates translation of abiD1 mRNA and consequently the AbiD1 phenotype, and occurs independently of phage infection. There is no synergism between the two abiD1 inducers. Purified Orf1 protein binds mRNAs containing a secondary structure motif, identified within the translation initiation regions of abiD1, the mid-infection phage bIL66 M-operon, and the L. lactis osmC gene.


CONCLUSIONS
Expression of the abiD1 gene and consequently AbiD1 phenotype is specifically translationally activated by the phage Orf1 protein. The loss of ability to activate translation of abiD1 mRNA determines the molecular basis for phage resistance to AbiD1. We show for the first time that temperature downshift also activates abortive infection by activation of abiD1 mRNA translation.
Bidnenko
Elena
E
Laboratoire de Génétique Microbienne, INRA, 78352 Jouy-en-Josas, France. elena.bidnenko@jouy.inra.fr
Chopin
Alain
A
Ehrlich
S Dusko
SD
Chopin
Marie-Christine
MC
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D001435:Bacteriophages / Q000235:genetics / Q000378:metabolism*; D003080:Cold Temperature; D015964:Gene Expression Regulation, Bacterial; D013294:Lactococcus lactis / Q000235:genetics* / Q000821:virology; D010641:Phenotype; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
C094328:AbiD protein, Lactococcus lactis; D001426:Bacterial Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins