Biology 457 & 557 -  Week 9

 

Activity of Transcription Factor E2F is Required for Entry into S Phase

 

many of the proteins necessary for DNA and deoxynucleotide synthesis are induced as

 

 

            the family of transcription factors that regulate transcription of these genes is called

  

 

 

1) RB protein

2) p107

 

           

            RB protein – its ability to inhibit the activity of E2F

 

           

                        ex: cultured cells derived from a tumor defective in both Rb alleles

 

 

                        if transfect these cells with a vector carrying Rb gene, what do you think

                        will happen?

 

 

 

 

 

                        bigger picture -  the mammalian G1 Cdk-cyclin complexes and Ckd2-cyclin A can

 

 

 

 

 

 

 

 

            in normal cycling cells, RB is

 

 

Role of Checkpoints in Cell Cycle Regulation

 

serious genetic damage can occur if cells progress to the next phase of the cell cycle

 

 

            1): induce S phase cells to enter mitosis (done by fusing these cells to cells

            in mitosis) because the MPF (maturation promoting factor – mentioned this

            when discussing oocyte maturation; initiation of mitosis in all euk. cells)

            present in the mitotic cell forces the chromosomes of the S phase cell to

            prematurely condense, which causes the chromosomes to fragment

 

            2): attachment of microtubules (spindle fibers) to kinetochores (centromeres)

            during metaphase, if not done correctly can result in

 

 

 

 

 

 

 

            3): cell can’t enter mitosis until all DNA is replicated

 

 

 

            4): defects in the assembly of mitotic spindle prevents exit from mitosis

 

                        have found that a checkpoint control somehow senses when the spindle

                        has not formed properly

 

 

 

                        MPF activity remains high, chromosomes remain condensed

 

 

                        this was described when yeast cells were treated with a microtubule-

                        depolymerizing drug called

 

                        at low conc. of benomyl, yeast cells took longer to assemble mitotic

                        spindle and attach kinetochores to microtubules (delay anaphase)

 

 

                        in contrast, mutants defective in this cell cycle checkpoint proceed through

                        anaphase before assembly of spindle

 

 

                        thought that at least one of the genes important to this is a

 

            5): DNA damage prevents entry into S phase and mitosis

 

                        if a cell is damaged by irradiation with UV light or by chemical treatment,

                        the cell arrests in G1

                        ex: G1 – the p53 tumor suppressor gene functions in the checkpoint

 

 

                        p53 codes for a transcription factor

 

 

 

                        cells are arrested in G1 until DNA damage is repaired

 

 

                        in a high % of human cancers

 

 

 

 

 

 

 

 

 

 

 

seen that cell cycle regulation is very important to induction of a cancerous cell……when lose control then the cell

 

 

 

 

think back to the lecture on cultured cells and I mentioned transformation as a mechanism whereby growth characteristics of cultured cells are altered (by viruses, chemicals, radiation) and they develop a tumor-forming capacity

 

what other events can trigger transformation?

 

            expression of one or a few

 

 

 

 

            oncology

 

 

 

 

 

 

ONCOGENES

 

characteristics:

 

of the many known oncogenes, almost all are derivatives of

 

these cellular counterparts are called

 

 

 

oncoproteins are then the protein products

 

 

a)     some oncoproteins are mutant forms of the protein product of

 

 

b)     sometimes the protein products of both the oncogene and proto-oncogene

are the same, but

 

 

                  in these cases, the mutation creating the oncogene changes

                 

 

a proto-oncogene and its related oncogene are designated with a 3 letter code

 

 

 

 

 

 

many oncogenes were first identified through the study of cancer induced by

 

 

            the retroviruses were found to contain or carry oncogenes

 

 

however, since not all retroviruses carry oncogenes, where did such genes originate?

 

            are they viral in origin?

            or might they represent genes captured by the virus from host animal cells?

 

 

 

 

the identification of the oncogene has frequently led to discovery of the corresponding

 

            the oncogene in a virus is denoted with a v prefix (ex: v-src)

           

 

oncogenes have also been identified by transfection

 

 

 

how many oncogenes have been discovered?

 

 

 

 

            5 types of proteins, depending on how they participate in control of cell growth

 

            oncogenes can code for :

            1)

            2)

            3)

            4)

            5)

 

 

 

 

 

 

 

growth factors:

            mentioned before that cells in culture can be induced to grow by addition of

            specific growth factors  ex: EGF -

                                                   ex: PDGF -

 

            these ligands serve as

           

            it is a way for cells to

 

            specific signals can produce a response in some cell types

 

            ex: EGF stimulates growth

 

            oncogenes rarely arise from genes

 

            really only one naturally occurring growth factor oncogene

            ex: sis

 

 

growth factor receptors:

a receptor binds a specific factor and then the receptor, recognizing it has bound this factor, will send a

 

            how do growth factor receptors work?

 

            these receptors have

 

            transmit the growth signal by:

 

            1) autophosphorylating

                        2) phophorylating

 

 

            normally, inactive receptors exist in an inactive

 

 

when bound by a growth factor, these receptors

 

 

            the activated receptor autophosphorylates additional

           

 

            these phosphorylated sites serve to recruit cytoplasmic targets of the

 

src gene 2

how then do these receptors become oncogenic?

 

1)     they are mutated in such a way as to keep the

 

           

2) gene amplification

 

            in most cases, much of the extracellular ligand-binding domain is deleted and the

            altered receptor has lost ligand control

             

           

                        ex: c-erb-B gene

 

            in another case, there is a single point mutation that results in an amino acid

substitution

            ex: neu oncogene

 

 

 

 

 

 

 

 

 

 

 

 

 

 

suggested that this substitution induces dimerization of the oncogenic Neu protein

 

 

 

or, the neu gene has been amplified and overexpressed

 

            this is found in human mammary carcinomas and ovarian cancers

 

also, it has been determined that amplification of the c-erb-B gene and subsequent

overexpression of the EGF receptor is common in

 

 

 

 

            a cell containing a constitutively active oncogenic receptor grows

            independently

           

 

            such mutant receptors can induce a spectrum of neoplasia