To define the actin-binding site within the NH2-terminal domain (residues 1-245) of chick smooth muscle alpha-actinin, we expressed a series of alpha-actinin deletion mutants in monkey Cos cells. Mutant alpha-actinins in which residues 2-19, 217-242, and 196-242 were deleted still retained the ability to target to actin filaments and filament ends, suggesting that the actin-binding site is loca...

Alpha-actinins are structural proteins of the Z-line. Human skeletal muscle expresses two alpha-actinin isoforms, alpha-actinin-2 and alpha-actinin-3, encoded by their respective genes ACTN2 and ACTN3. ACTN2 is expressed in all muscle fiber types, while only type II fibers, and particularly the type IIb fibers, express ACTN3. ACTN3 (R577X) polymorphism results in loss of alpha-actinin-3 and has...

During normal epithelial maturation, actin and α-actinin-4 accumulated at the cell junction over a period of several days (Fig. 1 A). By the second day post-confluence (2 dpc), most canonical junctional components, E-cadherin, α-catenin, and β-catenin, p120-catenin, and ZO-1, were already present (Fig. 1, A and B). However, vinculin has not been targeted at this early stage of junction developm...

In order to probe the organization of the contractile machinery in smooth muscle, we have studied the distribution of alpha-actinin, a protein present in high concentration in dense bodies, structures apparently analogous to the Z-disks of striated muscle. Localization of alpha-actinin in single isolated smooth muscle cells of the stomach muscularis of Bufo marinus was determined by analysis of...

The genome of the chlorarchiniophyte Bigelowiella natans codes for a protein annotated as an α-actinin-like protein. Analysis of the primary sequence indicate that this protein has the same domain structure as other α-actinins, a N-terminal actin-binding domain and a C-terminal calmodulin-like domain. These two domains are connected by a short rod domain, albeit long enough to form a single spe...

Immunoblotting studies with antisera against Z-protein, desmin, and alpha-actinin showed that Z-protein is clearly distinguishable from desmin and alpha-actinin. Z-protein is not a proteolytic product of another protein but is an intrinsic component of chicken breast muscle myofibrils. In these experiments, an SDS extract of intact muscle was first electrophoresed in a polyacrylamide gel, and t...

We used antibodies against alpha-actinin and myosin labeled directly with contrasting fluorochromes to localize these contractile proteins simultaneously in dividing chick embryo cells. During mitosis anti-alpha-actinin stains diffusely the entire cytoplasm including the mitotic spindle, while in the same cells intense antimyosin staining delineates the spindle. During cytokinesis both antibodi...

Recessive mutant gene c in axolotl embryos results in an absence of normal heart function. Immunofluorescence studies were done to determine the distributions of myosin, tropomyosin and alpha-actinin in the hearts of normal and mutant siblings. Anti-myosin specifically stains the A bands of myofibrils in normal hearts and reveals a progressive increase in myofibril organization with development...

............................................................................................................................................................. 9 REVIEW OF THE LITERATURE..................................................................................................................... 10 1. THE CELL DIVISION CYCLE AND CYCLIN-DEPENDENT KINASES........................................

A number of componenets of cytoskeleton are very large proteins which are in their complete native form non amenable to the structure determination via X-ray diffraction techniques due to their non-globular shape and intrinsic flexibility which hinder the formation of suitable crystals. Dissecting such proteins into their representative building blocks-domains and subsequent structural studies ...