Experimental shift in hosts' acceptance threshold of inaccurate-mimic brood parasite eggs

Hosts are expected to evolve resistance strategies that efficiently detect and resist exposure to virulent parasites and pathogens. When recognition is not error-proof, the acceptance threshold used by hosts to recognize parasites should be context dependent and become more restrictive with increasing predictability of parasitism. Here, we demonstrate that decisions of great reed warblers Acrocephalus arundinaceus to reject parasitism by the common cuckoo Cuculus canorus vary adaptively within a single egg-laying bout. Hosts typically accept one of their own eggs with experimentally added spots and the background colour left visible. In contrast, hosts reject such spotted eggs when individuals had been previously exposed to and rejected one of their own eggs whose background colour had been entirely masked. These results support patterns of adaptive modulation of antiparasitic strategies through shifts in the acceptance threshold of hosts and suggest a critical role for experience in the discrimination decisions between inaccurate-mimic parasite eggs and hosts' own eggs.     

An overview of the serpin superfamily

Serpins are a broadly distributed family of protease inhibitors that use a conformational change to inhibit target enzymes. They are central in controlling many important proteolytic cascades, including the mammalian coagulation pathways. Serpins are conformationally labile and many of the disease-linked mutations of serpins result in misfolding or in pathogenic, inactive polymers.     

Multimer Formation Is Not Essential for Nuclear Export of Human T-Cell Leukemia Virus Type 1 Rex trans-Activator Protein

The Rex trans-regulatory protein of human T-cell leukemia virus type 1 (HTLV-1) is required for the nuclear export of incompletely spliced and unspliced viral mRNAs and is therefore essential for virus replication. Rex is a nuclear phosphoprotein that directly binds to its cis-acting Rex response element RNA target sequence and constantly shuttles between the nucleus and cytoplasm. Moreover, Rex induces nuclear accumulation of unspliced viral RNA. Three protein domains which mediate nuclear import-RNA binding, nuclear export, and Rex oligomerization have been mapped within the 189-amino-acid Rex polypeptide. Here we identified a different region in the carboxy-terminal half of Rex which is also required for biological activity. In inactive mutants with mutations that map within this region, as well as in mutants that are deficient in Rex-specific multimerization, Rex trans activation could be reconstituted by fusion to a heterologous leucine zipper dimerization interface. The intracellular trafficking capabilities of wild-type and mutant Rex proteins reveal that biologically inactive and multimerization-deficient Rex mutants are still efficiently translocated from the nucleus to the cytoplasm. This observation indicates that multimerization is essential for Rex function but is not required for nuclear export. Finally, we are able to provide an improved model of the HTLV-1 Rex domain structure.     

Dominant-negative mutants are clustered in a domain of the human T-cell leukemia virus type I Rex protein: implications for trans dominance.

The 27-kDa Rex trans-acting protein appears to be essential for replication of human T-cell leukemia virus type I. Mutations introduced outside of the Rex RNA-binding domain-nucleolar localization signal display either wild-type activity or, conversely, yield dominant-negative proteins. We generated missense mutations in a particular domain of the Rex protein (amino acid residues 54 to 69) which is characterized by a cluster of dominant-negative mutants. Our results indicate that amino acids 57 to 67 are critically important for Rex function mediated through the RxRE cis-acting RNA sequence. Within this domain, only amino acids 61 to 63 could be mutated without loss of function. All other missense and deletion mutants yielded dominant-negative proteins. In vitro RNA-binding studies performed with glutathione S-transferase-Rex fusion proteins demonstrated that all of the mutant Rex proteins interacted specifically with RxRE RNA. Analysis of chimeric Rex-Rev proteins suggests that this Rex domain is important for oligomerization.     

Bateman's Principle in Cooperatively Breeding Vertebrates: The Effects of Non-breeding Alloparents on Variability in Female and Male Reproductive Success

The sex-specific slopes of Bateman's gradients have importantimplications for understanding animal mating systems, includingpatterns of sexual selection and reproductive competition. Intersexualdifferences in the fitness benefits derived from mating withmultiple partners are expected to yield distinct patterns ofreproductive success for males and females, with variance indirect fitness predicted to be greater among males. These analysesassume that typically all adults are reproductive and that failureto produce offspring is non-adaptive. Among some species ofcooperatively breeding birds and mammals, however, non-breedingadult alloparents are common and may comprise the majority ofindividuals in social groups. The presence of a large numberof non-breeding adults, particularly when coupled with greatersocial suppression of reproduction among females, may alterthe relative variance in direct fitness between the sexes, therebygenerating an apparent contradiction to Bateman's Paradigm.To explore quantitatively the effects of non-breeding alloparentson variance in reproductive success, we used genetic estimatesof parentage and reproductive success drawn from the literatureto calculate the relative variability in direct fitness forfemales and males in alloparental and "other" societies of birdsand mammals. Our analyses indicate that in mammals and, to alesser extent, in birds, variability in direct fitness is greateramong females in species characterized by the presence of non-breedingalloparents. These data suggest that social interactions, includingsocial suppression of reproduction, are powerful determinantsof individual direct fitness that may modify sex-specific patternsof reproductive variance from those described by Bateman.     

A password for species recognition in a brood-parasitic bird

Recognition of conspecifics is an essential precursor of sexual reproduction. Most mammals and birds learn salient features of their parents or siblings early in ontogeny and later recognize individuals whose phenotypes match the mental image (template) of relatives closely enough as conspecifics. However, the young of brood parasites are reared among heterospecifics, so social learning will yield inappropriate species recognition templates. Initially, it was inferred that conspecific recognition in brood parasites depended on genetically determined templates. More recently it was demonstrated that learning plays a critical role in the development of parasites' social preferences. Here we propose a mechanism that accommodates the interaction of learned and genetic components of recognition. We suggest that conspecific recognition is initiated when a young parasite encounters some unique species-specific signal or "password" (e.g. a vocalization, behaviour or other characteristic) that triggers learning of additional aspects of the password-giver's phenotype. We examined the possibility that nestlings of the obligately brood-parasitic brown-headed cowbird (Molothrus ater) could use a species-specific vocalization, the "chatter", as a password. We found that six-day-old nestlings responded (begged) significantly more frequently to playbacks of chatters than to other avian sounds and that two-month-old fledglings approached playbacks of chatters more quickly than vocalizations of heterospecifics. Free-living cowbird fledglings and adults also approached playbacks of chatters more often than control sounds. Passwords may be involved in the ontogeny of species recognition in brood parasites generally.     

Direct observation of nucleocytoplasmic transport by microinjection of GFP-tagged proteins in living cells.

We established a straightforward experimental system to investigate directly the requirements for nucleocytoplasmic transport in live cells. For this purpose, substrates were created containing nuclear localization signals (NLS) or nuclear export signals (NES) linked to a chimeric protein composed of the glutathione S-transferase (GST) fused to the green fluorescent protein (GFP). The combination of GST/GFP-tagging allowed us to control protein expression in bacteria and to monitor protein purification during chromatography. Following microinjection into somatic cells, nuclear export/import of the highly fluorescent substrates could be observed directly by fluorescence microscopy. This system sets the stage to quantitate, in real time, the kinetics of nuclear import/export in living cells and to evaluate qualitative differences in various NLS/NES signals and pathways.     

A novel mouse model for inhibition of DOHH-mediated hypusine modification reveals a crucial function in embryonic development,proliferation and oncogenic transformation

The central importance of translational control by post-translational modification has spurred major interest in regulatory pathways that control translation. One such pathway uniquely adds hypusine to eukaryotic initiation factor 5A (eIF5A), and thereby affects protein synthesis and, subsequently, cellular proliferation through an unknown mechanism. Using a novel conditional knockout mouse model and a Caenorhabditis elegans knockout model, we found an evolutionarily conserved role for the DOHH-mediated second step of hypusine synthesis in early embryonic development. At the cellular level, we observed reduced proliferation and induction of senescence in 3T3 Dohh^−/− cells as well as reduced capability for malignant transformation. Furthermore, mass spectrometry showed that deletion of DOHH results in an unexpected complete loss of hypusine modification. Our results provide new biological insight into the physiological roles of the second step of the hypusination of eIF5A. Moreover, the conditional mouse model presented here provides a powerful tool for manipulating hypusine modification in a temporal and spatial manner, to analyse both how this unique modification normally functions in vivo as well as how it contributes to different pathological conditions.KEY WORDS: Hypusine modification, Translational control, Cancer, Mouse models