Retroviral mediated expression of the human myeloid nuclear antigen in a null cell line upregulates Dlk1 expression

J Cell Biochem. 2002;86(1):56-66. doi: 10.1002/jcb.10190.

Abstract

The human myeloid nuclear differentiation antigen (MNDA) is a hematopoietic cell specific nuclear protein. MNDA and other related gene products interact with and alter the activity of a large number of proteins involved in regulating specific gene transcription. MNDA and related genes exhibit expression characteristics, which suggest functions unique to specific lineages of cells, in addition to mediating the effects of interferons. Cells of the human K562 myeloid line do not express MNDA and are relatively immature compared to lines that express MNDA (HL-60, U937, and THP1). The hypothesis that MNDA influences the expression of specific genes was tested by creating MNDA expressing K562 cells using stable retroviral mediated gene transfer followed by evaluation of transcription profiles. Two macroarrays containing a total of 2,350 cDNAs of known genes showed a specific up-regulation of Dlk1 expression in MNDA expressing K562 cell clones. Real time quantitative RT-PCR analysis confirmed an average of over 3- and 7-fold upregulation of Dlk1 in two clones of MNDA expressing K562 cells. The effects on Dlk1 were also confirmed by Northern blotting. Dlk1 is essential for normal hematopoiesis and abnormal expression is a proposed marker of myelodysplastic syndrome. Additional screening of transcription profiles after induced erythroid and megakaryoblastic differentiation showed no additional gene transcripts altered by the presence of MNDA. These results indicate that MNDA alters expression of a gene essential for normal hematopoiesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Differentiation, Myelomonocytic / genetics*
  • Antigens, Differentiation, Myelomonocytic / metabolism*
  • Cell Differentiation / genetics
  • Epidermal Growth Factor / genetics
  • Epidermal Growth Factor / metabolism*
  • Gene Deletion
  • Gene Expression*
  • Genetic Vectors / genetics
  • Humans
  • K562 Cells
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Mice
  • Oligonucleotide Array Sequence Analysis
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Retroviridae / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism*
  • Transcription, Genetic
  • Up-Regulation

Substances

  • Antigens, Differentiation, Myelomonocytic
  • MNDA protein, human
  • Membrane Proteins
  • RNA, Messenger
  • Transcription Factors
  • Epidermal Growth Factor