Redistribution of motor proteins by Cytochalasin J treatment

Robinson, Rollin W.; Snyder, Judith A.

doi:10.1016/s1065-6995(03)00123-9

Cited by 3 publications

(3 citation statements)

References 42 publications

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“…Cytochalasin J is not a particularly potent cytochalasin (Cooper, 1987;Yarhara et al, 1982) yet it has the most significant effect on PtK 1 cells mitotic events when compared with similar concentrations of CB, CD, CE and CH (unpublished data). Cytochalasin J treatment of mitotic PtK 1 cells indicates its effects not only the microfilament (MF) system, but also the dynein complex (Robinson and Snyder, 2003). Since dynactin contains Arp1 and directly attaches to p150 Glued , it may also be sensitive to CJ treatment, particularly since the ''actin fold'' has a high homology with the ATP binding site with actin filaments (Kabsch and Holmes, 1995).…”

Section: Discussionmentioning

confidence: 99%

Colocalization studies of Arp1 and p150Glued to spindle microtubules during mitosis: The effect of cytochalasin on the organization of microtubules and motor proteins in PtK1 cells

Robinson¹,

Snyder²

2006

Cell Biology International

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Motor proteins play a fundamental role in the congression and segregation of chromosomes in mitosis as well as the formation of the mitotic spindle. In particular, the dynein/dynactin complex is involved in the maintenance of the spindle, formation of astral microtubules, chromosome motion, and chromosome segregation. Dynactin is a multisubunit, high molecular weight protein that is responsible for the attachment of cargo to dynein. There are a number of major subunits in dynactin that are presumed to be important during mitosis. Arp1 is thought to be the attachment site for cargo to the complex while p150(Glued), a side arm of this complex regulates binding to MTs and the binding of dynactin to dynein. We performed colocalization studies of Arp1 and p150(Glued) to spindle microtubules. Both Arp1 and p150(Glued) colocalize with spindle MTs as well as cytoplasmic components. When treated with cytochalasin J, Arp1 concentrates at the centrosomes and is less co-localized with spindle MTs. Cytochalasin J has less of an effect on the colocalization of p150(Glued) with spindle MTs, suggesting that Arp1 may have a cytochalasin J sensitive site.

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Section: Discussionmentioning

confidence: 99%

Colocalization studies of Arp1 and p150Glued to spindle microtubules during mitosis: The effect of cytochalasin on the organization of microtubules and motor proteins in PtK1 cells

Robinson¹,

Snyder²

2006

Cell Biology International

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“…Interphase cells revealed a localization of MTs emanating from the centrosome near the nucleus ( Fig. 1A; Robinson and Snyder 2003). However, when cells were fixed with PFA the MT array appeared fragmented with shorter and fewer MTs (Fig.…”

Section: Microtubules With Dapimentioning

confidence: 92%

An innovative fixative for cytoskeletal components allows high resolution in colocalization studies using immunofluorescence techniques

Robinson

Snyder

2003

Histochemistry and Cell Biology

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Using a new fixation solution, CytoSkelFix, it is now possible to obtain superior fixation and thus resolution of cytoskeletal components using immunofluorescence and fluorescence microscopy. This fixative combines rapid cell penetration and cellular crosslinking of proteins such that both preservation and resolution of cellular proteins can be detected. The cytoskeleton has proven very difficult to preserve, partly because of the lability of one of the filament systems (microtubules), and one single fixative is incapable of properly preserving microtubules, microfilaments, and intermediate filaments for localization in the same cell. Further, the motor proteins associated with the cytoskeletal elements are even more difficult to preserve, particularly simultaneously with the fiber system with which they associate. We present evidence that CytoSkelFix is a superior preservative and would be useful in fixation for all types of immunofluorescence colocalization studies where superior preservation is required.

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“…Most hypotheses of force production focus solely on spindle microtubules and their motor proteins (Gorbsky et al, 1988;Nicklas, 1989;Desai et al, 1998;Sharp et el., 2000;LaFountain et al, 2004;Mitchison, 2005;Rogers et al, 2005;Rath et al, 2009). Others have considered components such as actin (Forer and Pickett-Heaps, 1998;Silverman-Gavrila and Forer, 2000;Forer, et al, 2003;Robinson and Snyder, 2003;Snyder et al, 2010), myosin 2005Forer, 2000, 2003;Rosenblatt et al, 2004;Woolner et al, 2008;Snyder et al, 2010;Rump et al, 2011), titin and spindle matrix proteins (Johansen and Johansen, 2007;Lince-Faria et al, 2009;Qi et al, 2009;Ding et al, 2009;Johansen et al, 2011 ). This article describes experiments that deal with the role of spindle myosin in anaphase chromosome movement.…”

Section: Introductionmentioning

confidence: 99%

The role of myosin phosphorylation in anaphase chromosome movement

Sheykhani¹,

Shirodkar²,

Forer³

2013

European Journal of Cell Biology

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This work deals with the role of myosin phosphorylation in anaphase chromosome movement. Y27632 and ML7 block two different pathways for phosphorylation of the myosin regulatory light chain (MRLC). Both stopped or slowed chromosome movement when added to anaphase crane-fly spermatocytes. To confirm that the effects of the pharmacological agents were on the presumed targets, we studied cells stained with antibodies against mono- or bi-phosphorylated myosin. For all chromosomes whose movements were affected by a drug, the corresponding spindle fibres of the affected chromosomes had reduced levels of 1P- and 2P-myosin. Thus the drugs acted on the presumed target and myosin phosphorylation is involved in anaphase force production. Calyculin A, an inhibitor of MRLC dephosphorylation, reversed and accelerated the altered movements caused by Y27632 and ML-7, suggesting that another phosphorylation pathway is involved in phosphorylation of spindle myosin. Staurosporine, a more general phosphorylation inhibitor, also reduced the levels of MRLC phosphorylation and caused anaphase chromosomes to stop or slow. The effects of staurosporine on chromosome movements were not reversed by Calyculin A, confirming that another phosphorylation pathway is involved in phosphorylation of spindle myosin.

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Redistribution of motor proteins by Cytochalasin J treatment

Cited by 3 publications

References 42 publications

Colocalization studies of Arp1 and p150Glued to spindle microtubules during mitosis: The effect of cytochalasin on the organization of microtubules and motor proteins in PtK1 cells

Colocalization studies of Arp1 and p150Glued to spindle microtubules during mitosis: The effect of cytochalasin on the organization of microtubules and motor proteins in PtK1 cells

An innovative fixative for cytoskeletal components allows high resolution in colocalization studies using immunofluorescence techniques

The role of myosin phosphorylation in anaphase chromosome movement

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