Ab. Fulton et C. Alftine, ORGANIZATION OF PROTEIN AND MESSENGER-RNA FOR TITIN AND OTHER MYOFIBRIL COMPONENTS DURING MYOFIBRILLOGENESIS IN CULTURED CHICKEN SKELETAL-MUSCLE, Cell structure and function, 22(1), 1997, pp. 51-58
Myofibril assembly requires the cell to join diverse components, corre
ctly oriented to the rest of the cell. Titin, a huge elastic protein w
ith a role in myogenesis, assembles during translation in vivo and may
require spatially organized mRNA to allow assembly. By immunofluoresc
ence, we examined titin and myosin protein organization early in skele
tal muscle development in vitro; titin was the first organized, initia
lly as spots, then as periodically spaced lines, and later as doublets
. Titin mRNA organization during development was detected by fluoresce
nt in situ hybridization. Only titin mRNA was seen in mononucleated my
oblasts. Shortly after fusion, both titin protein and mRNA were diffus
e. Titin mRNA remained diffuse when titin protein formed cables. Where
titin protein formed linear arrays of spots, titin mRNA showed a coli
near, continuous array. Titin mRNA remained in arrays colinear with yo
ung myofibrils until several slender myofibrils aligned laterally; the
n, titin mRNA formed periodic arrays. The titin probe encodes peptide
sequence in the A band, where this region of titin mRNA is detected in
the most organized cells. Nebulin undergoes a similar progression sli
ghtly later in development. This pattern, of narrowly spaced stripes,
is too closely spaced to function in the soluble phase. Titin mRNA is
the earliest mRNA to become so highly organized in muscle; it does so
earlier and at a different location than do mRNAs for costamere protei
ns. These results, taken with earlier ones, suggest mRNA localization
may be as key to somatic cell differentiation as it is to embryonic de
velopment.