Transcriptomics of a KDELR1 knockout cell line reveals modulated cell adhesion properties
Transcriptomics of a KDELR1 knockout cell line reveals modulated cell adhesion properties
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DOI：
10.1038/s41598-019-47027-5
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发表时间：
2019-07-23
影响因子：
4.6
通讯作者：
Schmitt, Manfred J.
中科院分区：
文献类型：
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作者：
Blum, Andrea;Khalifa, Saleem;Schmitt, Manfred J.
KDEL受体（KDELRs）代表分泌途径的跨膜蛋白，其调节可溶性ER-居民的保留以及逆行和顺行囊泡运输。此外，KDELRs参与细胞应激反应和ECM降解的调节。为了更深入地了解KDELR 1特异性功能，我们通过比较KDELR 1-KO细胞与其各自的HAP 1野生型，通过全转录组分析表征了KDELR 1-KO细胞系（HAP 1）。我们的数据表明，超过300个显著和差异表达的基因，其基因产物主要参与发育过程，如细胞粘附和ECM组成，指出由于KDELR 1的丢失导致严重的细胞疾病。通过体外粘附测定进一步证明了KDELR 1-KO细胞的粘附能力受损，而胶原蛋白和/或层粘连蛋白包被使KDELR 1-KO细胞的粘附性质与野生型相比几乎加倍，证实了改善或恢复细胞粘附能力的转录适应。在ER应激条件下，ER驻留PDI的分泌增加和细胞活力降低证实了分泌途径内的扰动，表明KDELR 1-KO细胞在维持细胞稳态方面严重受损。
KDEL receptors (KDELRs) represent transmembrane proteins of the secretory pathway which regulate the retention of soluble ER-residents as well as retrograde and anterograde vesicle trafficking. In addition, KDELRs are involved in the regulation of cellular stress response and ECM degradation. For a deeper insight into KDELR1 specific functions, we characterised a KDELR1-KO cell line (HAP1) through whole transcriptome analysis by comparing KDELR1-KO cells with its respective HAP1 wild-type. Our data indicate more than 300 significantly and differentially expressed genes whose gene products are mainly involved in developmental processes such as cell adhesion and ECM composition, pointing out to severe cellular disorders due to a loss of KDELR1. Impaired adhesion capacity of KDELR1-KO cells was further demonstrated through in vitro adhesion assays, while collagen- and/or laminin-coating nearly doubled the adhesion property of KDELR1-KO cells compared to wild-type, confirming a transcriptional adaptation to improve or restore the cellular adhesion capability. Perturbations within the secretory pathway were verified by an increased secretion of ER-resident PDI and decreased cell viability under ER stress conditions, suggesting KDELR1-KO cells to be severely impaired in maintaining cellular homeostasis.