RESUMO
LMO7 (LIM domain only 7) is a transcription regulator for expression of many Emery-Dreifuss muscular dystrophy-relevant genes, and binds to α-actinin and AF6/afadin at adherens junctions for epithelial cell-cell adhesion. In this study, we found that human LMO7 interacted with the spindle assembly checkpoint (SAC) protein MAD1. LMO7 colocalized with actin filaments at the cell membrane but did not colocalize with MAD1 at kinetochores in prometaphase. Our observations reveal that overexpression but not depletion of LMO7 caused a SAC defect, and that the LIM domain of LMO7 was a determinant of its ability to interfere with kinetochore localization of the SAC proteins MAD2 and BUBR1 and cause a SAC defect though the LIM peptide itself did neither bind to MAD1, MAD2 and BUBR1 nor localize to the actin filaments. However, overexpression of LMO7 or the LIM peptide did not interfere with kinetochore localization of MAD1. Additionally, overexpression of the LIM peptide prolonged mitotic timing and interfered with chromosome congression whereas that of LMO7b did not. Taken together, we conclude that LMO7 via its LIM domain acts to control mitosis progression and exerts an effect on the SAC.
Assuntos
Citoesqueleto de Actina/metabolismo , Proteínas de Ciclo Celular/metabolismo , Membrana Celular/metabolismo , Proteínas com Domínio LIM/metabolismo , Pontos de Checagem da Fase M do Ciclo Celular , Mitose , Proteínas Nucleares/metabolismo , Fatores de Transcrição/metabolismo , Animais , Proteínas de Ciclo Celular/química , Proteínas de Ciclo Celular/genética , Linhagem Celular Tumoral , Humanos , Interfase , Cinetocoros/metabolismo , Proteínas com Domínio LIM/antagonistas & inibidores , Proteínas com Domínio LIM/química , Proteínas com Domínio LIM/genética , Proteínas Luminescentes/química , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Metáfase , Proteínas Nucleares/química , Proteínas Nucleares/genética , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/metabolismo , Prometáfase , Domínios Proteicos , Multimerização Proteica , Transporte Proteico , Interferência de RNA , Ratos , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/metabolismo , Polos do Fuso/metabolismo , Fatores de Transcrição/antagonistas & inibidores , Fatores de Transcrição/química , Fatores de Transcrição/genéticaRESUMO
This study aimed to compare and analyze the changes in the coagulation factors in fresh frozen plasma (FFP) prior to and following leukocyte filtration and irradiation. In total, 30 bags of FFP from healthy donors were processed: One-third of the FFP of each bag was left within the original bag (the A group), the other two-thirds of the FFP of each bag were passed through a disposable leukocyte filter, then divided equally into two parts. One of these was designated as the B group, and the other was designated the C group (subjected to 30 Gy irradiation). All samples were analyzed to evaluate 16 coagulation indicators. Analysis of variance revealed that there were statistically significant differences in the levels of fibrinogen (FbgC) and coagulation factor VIII (FVIII:C) among the groups (P=0.044 and P=0.015, respectively); the Dunnett's t-test revealed that there was a statistically significant difference in the level of FbgC between the A and B groups (P=0.025), and there was a statistically significant difference in the level of FVIII:C between the A and C groups (P=0.009); while the remaining 14 coagulation parameters were not significantly different among the groups. Although the levels of FbgC and FVIII:C in the FFP were reduced following treatment, this would not affect the clinical effect of the FFP.