Intracellular factors are involved in and essential for hematopoiesis. HIV-1 Tat-interacting protein of 110 kDa (TIP110; p110 nrb / SART3/p110) is an RNA-binding nuclear protein implicated in the regulation of HIV-1 gene and host gene transcription, premRNA splicing, and cancer immunology. In the present study, we demonstrate a role for TIP110 in the regulation of hematopoiesis. TIP110 was expressed in human CD34 ؉ cells and decreased with differentiation of CD34 ؉ cells. TIP110 mRNA was also expressed in phenotyped mouse marrow hematopoietic stem cells (HSCs) and hematopoietic progenitor cells (HPCs). Using TIP110 transgenic (TIP110 TG ) and haploinsufficient (TIP110 ؉/؊ ) mice, we found that increased TIP110 expression enhanced HPC numbers, survival, and cell cycling, whereas decreased TIP110 expression had the opposite effects. Moreover, TIP110 ؉/؊ bone marrow HPCs responded more effectively, and TIP110 TG HPCs less effectively, than those of wild-type control mice to recovery from the cell-cycle-active drug 5-fluorouracil (5-FU). Unexplained sex differences were noted in HSC competitive repopulating ability, but not HPC numbers, in TIP110 TG mice. Intracellularly, TIP110 regulated CMYC and GATA2 expression at the transcriptional level, and TIP110 and CMYC reciprocally regulated the expression of each other. These results demonstrate a role for TIP110 in the regulation of hematopoiesis, effects that are likely linked to TIP110 regulation of CMYC.
IntroductionHematopoiesis is regulated at the level of hematopoietic stem cells (HSCs) and hematopoietic progenitor cells (HPCs) via intracellular and extracellular molecules. [1][2][3] Homeostasis is crucial for normal hematopoiesis. 2 Cell cycling of HSCs/HPCs is regulated by growth factors/cytokines, as well as by transcription factors such as C-MYB, CMYC, GATA2, HOX family proteins, and BMI-1. C-MYB promotes HSC growth through induced CMYC. [4][5][6] CMYC induces the expression of positive regulators involved in G1/S cell-cycle transition, and also represses negative regulators of the cell cycle, events required for the proliferation of hematopoietic progenitors. 7 CMYC is involved in cell proliferation and growth, angiogenesis, apoptosis, differentiation, and regulation of chromatin structure, [8][9][10][11] and plays a role in controlling self-renewal and differentiation of HSCs. 12 Tat-interacting protein of 110 kDa (TIP110), which was initially cloned from a human KG-1 myeloid cell line cDNA library, 13 is implicated in RNA metabolism and tumor-antigen presentation. [14][15][16][17] We demonstrated previously that TIP110 regulates the transcription of HIV-1 and cellular genes. 18,19 It is considered a mammalian homolog of the yeast Prp24 protein, and functions as a general pre-mRNA splicing factor. 20 Using molecular, cellular, and genetic strategies, including TIP110 transgenic (TIP110 TG ) and conditional knockout (TIP110 ϩ/Ϫ ) mice, ectopic expression, and shRNA knockdown, we found that TIP110 regulates HPC/HSC function, effects likely involving cell-cycl...