Proteomics of hyposaline stress in blue mussel congeners (genus Mytilus): implications for biogeographic range limits in response to climate change

Climate change is affecting species' physiology, pushing environmental tolerance limits and shifting distribution ranges. In addition to temperature and ocean acidification, increasing levels of hyposaline stress due to extreme precipitation events and freshwater runoff may be driving some of the reported recent range shifts in marine organisms. Using two-dimensional gel electrophoresis and tandem mass spectrometry, we characterized the proteomic responses of the cold-adapted blue mussel Mytilus trossulus , a native to the Pacific coast of North America, and the warm-adapted M. galloprovincialis , a Mediterranean invader that has replaced the native from the southern part of its range, but may be limited from expanding north due to hyposaline stress. After exposing laboratory-acclimated mussels for 4 h to two different experimental treatments of hyposaline conditions and one control treatment (24.5, 29.8 and 35.0 psu, respectively) followed by a 0 and 24 h recovery at ambient salinity (35 psu), we detected changes in the abundance of molecular chaperones of the endoplasmic reticulum (ER), indicating protein unfolding, during stress exposure. Other common responses included changes in small GTPases of the Ras superfamily during recovery, which suggests a role for vesicle transport, and cytoskeletal adjustments associated with cell volume, as indicated by cytoskeletal elements such as actin, tubulin, intermediate filaments and several actin-binding regulatory proteins. Changes of proteins involved in energy metabolism and scavenging of reactive oxygen species suggest a reduction in overall energy metabolism during recovery. Principal component analyses of protein abundances suggest that M. trossulus is able to respond to a greater hyposaline challenge (24.5 psu) than M. galloprovincialis (29.8 psu), as shown by changing abundances of proteins involved in protein chaperoning, vesicle transport, cytoskeletal adjustments by actin-regulatory proteins, energy metabolism and oxidative stress. While proteins involved in energy metabolism were lower in M. trossulus during recovery from hyposaline stress, M. galloprovincialis showed higher abundances of those proteins at 29.8 psu, suggesting an energetic constraint in the invader but not the native congener. Both species showed lower levels of oxidative stress proteins during recovery. In addition, oxidative stress proteins associated with protein synthesis and folding in the ER showed lower levels during recovery in M. galloprovincialis , in parallel with ER chaperones, indicating a reduction in protein synthesis. These differences may enable the native M. trossulus to cope with greater hyposaline stress in the northern part of its range, as well as to outcompete M. galloprovincialis in the southern part of M. trossulus ' range, thereby preventing M. galloprovincialis from expanding further north. * ALDH : aldehyde dehydrogenase BiP : binding immunoglobulin protein CCT : chaperonin containing TCP-1 DLDH : dihydrolipoyl dehydrogenase ER : endoplasmic reticulum EST : expressed sequence tag ETC : electron transport chain FABP : fatty-acid binding protein F(G)-actin : filamentous (globular)-actin FADH2 : flavin adenine dinucleotide dihydrogen GRP : glucose-regulated protein G(D)TP : guanosine 5’-(di-)triphosphate GCA : M. galloprovincialis ; cytoskeleton-associated proteins; cluster A GCHA : M. galloprovincialis ; protein chaperoning/degradation; cluster A GEA : M. galloprovincialis ; energy metabolism; cluster A HSC : heat shock cognate HSP : heat shock protein ICDH : isocitrate dehydrogenase IPG : immobilized pH gradient MOWSE : molecular weight search NAD(H) : nicotinamide adenine dinucleotide (reduced form) NADP(H) : nicotinamide adenine dinucleotide phosphate (reduced form) NHE-RF : Na+/H+ exchange regulatory factor PC : principal component PCA : principal component analysis PDH : pyruvate dehydrogenase PDI : protein disulfide isomerase PMF : peptide mass fingerprint PTM : post-translational modification Rab-GDI : Rat Brain (small GTPase)-GDP dissociation inhibitor Ras : Rat-sarcoma (small GTPase) Rho : Ras-homology (small GTPase) ROS : reactive oxygen species RSH : radial spoke head SAR : Secretion-associated Ras-like (small GTPase) Sec61 : ER protein transport protein SOD : superoxide dismutase TCP-1 : T-complex protein 1 TCA : M. trossulus ; cytoskeleton-associated proteins; cluster A TCHA : M. trossulus ; protein chaperoning/degradation; cluster A TEA : M. trossulus ; energy metabolism; cluster A