The proteins produced just prior to maturation desiccation in the developing, orthodox seed, are stored in the desiccated state and recruited as the functional proteome upon imbibition. For the resumption of protein function, these stored proteins must be protected from permanent denaturation while dehydrating, throughout desiccation, and during rehydration. For some forms of damage there is the possibility of repair following imbibition potentially coordinated with de-aggregation into monodispersed polypeptides capable of refolding into a functional configuration. While studying aspects of the natural protection and repair mechanism in seeds, evidence has accrued that those proteins directly involved in translation are particular targets of both protection and protein repair. Such a phenomenon was first described by Rajjou et al. (2008) examining the frequency with which proteins involved in translation were identified as differentially abundant between aged and un-aged Arabidopsis seeds and the translational competence of aged versus un-aged seeds. The inference drawn from these observations was that, of all the stored proteins, it is imperative that those involved in translation endure desiccation, quiescence and rehydration in a functional state if the seed is to survive. Proteins involved in any other process other than translation can be replaced from the stored transcriptome or by de novo transcription but no mRNA is of value without the translational machinery. This has become known as ‘Job's rule’ in honour of the laboratory from which this hypothesis was first put forward (Rajjou et al., 2008). We review in this manuscript the evidence accrued to date on which Job's rule is based.

Escherichia coli ribosomal protein L4 autogenously regulates transcription of the S10 operon, which encodes L4 and 10 other ribosomal proteins. Regulation results from L4-stimulated premature transcription termination at a U-rich site in the untranslated leader. The process requires transcription factor NusA. Here we report a detailed analysis of the RNA requirements for NusA-dependent, L4-mediated transcription control. We found that efficient regulation requires multiple features of the S10 leader, including two hairpins, called HD and upper HE, a connecting tether, and a U-rich sequence at the distal side of HE. As expected, regulation was optimal when all 7 Us were maintained in the U4CGU3 sequence at the termination site. However, despite the apparent specificity of L4 action on only the S10 operon, there is surprising flexibility at the primary sequence level for the HD-tether-HE region. Changes in the sequence of non-base-paired nucleotides flanking the HD hairpin or an A at the second position of the HD loop reduced L4 regulation, but other changes had little or no effect. Furthermore, generic hairpins from other RNAs could replace the natural HD and upper HE hairpins with little or no reduction of L4 control, suggesting that the secondary structure elements are also relatively generic. The lack of specific sequence requirements suggests that L4 may recognize multiple elements within this region of the nascent leader.

The transcript of the Saccharomyces cerevisiae gene, RPL30, is subject to regulated splicing and regulated translation, due to a structure that interacts with its own product, ribosomal protein L30. We have followed the fate of the regulated RPL30 transcripts in vivo. Initially, these transcripts abortively enter the splicing pathway, forming an unusually stable association with U1 snRNP. A large proportion of the unspliced molecules, however, are found in the cytoplasm. Most of these are still bound by L30, as only a small fraction are engaged in translation. Eventually, the unspliced RPL30 transcripts escape the grasp of L30, associate with ribosomes, and fall prey to nonsense mediated decay.

Murine antibodies, raised against a purified recombinant 30 kDa laminin-binding protein from Echinococcus granulosus, were used to investigate the tissue distribution of the native protein in protoscoleces and brood capsules. Immunofluorescence, in combination with confocal microscopy, revealed that the protein was distributed in small annular foci near the peripheral regions of the protoscoleces. Immunoelectron microscopy of thawed cryosections demonstrated that the laminin-binding protein was present in the cytoplasm of tegumentary cytons and myocytons, but not in cells of the excretory system. The protein was associated with amorphous regions of the cytoplasm, and was not expressed at the surfaces of cells. This distribution resembles those of other invertebrate laminin-binding proteins, which are thought to act in the cell cycle and cell proliferation events. A low degree of label was consistently detected in extracellular matrices of the protoscolex.