Comparison of the variable loop regions of myosin heavy chain genes from Antarctic and temperate isopods.
The evolutionary adaptations of functional genes to life at low temperatures are not well characterised in marine and fresh water invertebrates. Temperature has been shown to affect the functional characteristics of fish muscles, with changes in the velocity of shortening and ATPase activity being associated with myosin heavy chain (MyHC) isoform composition and the structure of the surface loop regions. Two PCR products spanning loops 1 and 2 of a MyHC gene from an Antarctic isopod (Glyptonotus antarcticus) were sequenced and compared with those of a temperate isopod (Idotea resecata), slow and fast fibres from lobster (Homarus gammarus) and a cold water amphipod (Eulimnogammarus verrucosus), revealing specific differences between the species, possibly related to fibre type and habitat temperature. The loop 2 region from G. antarcticus myosin was cloned and used for Northern analysis of total RNA from the other species. The cloned myosin cDNA hybridised specifically to a 6.6-kb transcript, in G. antarcticus muscle. In contrast, cDNA probes for lobster slow myosin and actin hybridised to muscle RNA from all species, demonstrating that a distinct MyHC isoform is expressed in the Antarctic isopod, as opposed to the temperate species. The inter- and intra-specific sequence differences in loop 2 region suggest that this may be a site for muscle adaptation to enable function at the low temperatures found in the Southern Ocean.
Holmes
J M
JM
Centre for Science and Technology, School of Postgraduate Medicine, Keele University, Thornburrow Drive, North Staffordshire ST4 7QB, Stoke on Trent, UK. holmes.janet@hotmail.com
Whiteley
N M
NM
Magnay
J L
JL
El Haj
A J
AJ
D000199:Actins / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000864:Antarctic Regions; D015152:Blotting, Northern; D003445:Crustacea / Q000235:genetics*; D017931:DNA Primers / Q000737:chemistry; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000737:chemistry*; D018995:Myosin Heavy Chains / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D009218:Myosins / Q000235:genetics / Q000378:metabolism; D008121:Nephropidae / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D000199:Actins; D017931:DNA Primers; D012313:RNA; D018995:Myosin Heavy Chains; D009218:Myosins