Why would constitutively active Ras

or Src lead to cancer ?

How do Ras and Src act in our body, in vivo ?

From cell culture to model organisms

 So, what job does
the protein encoded by src
do within the cell?

The first BIG step:

using serum to
immunoprecipitate the
v-Src protein

This led to the discovery that Src

is post-translationally modified
 Protein kinases and protein phosphatases
add and remove phosphate groups from
target proteins

Lodish et al. Fig. 20-5

Figure 5.5a The Biology of Cancer (© Garland Science 2007)
Which amino acids can be phosphorylated?

And Why those amino acids??

 Src is a Tyrosine Kinase
As a kinase, it can affect many cellular events

Figure 15-18a Molecular Biology of the Cell (© Garland Science 2008)

Normally, Src kinase intrinsic activity is low
What makes Src so active in transformed cells?

Western Blot with antibody

that recognizes
Tyr phosphorylated
proteins
 The structures of c-src and v-src
provided an important clue!

Lodish et al. Fig. 24-17

 Src contains three domains
that are shared with other proteins
Binds polyproline motifs Phosphorylates
other proteins

Binds peptides phosphorylated on Tyr

Scientists have determined
the precise 3-dimensional structure of Src

Xu et al. Nature. 1997 385:595-602

 Tyrosine phosphorylation of the C-terminus
creates an intramolecular
and inhibitory interaction

Lodish et al. Fig. 24-17

Src is normally
inactive due to
intramolecular
inhibition

Lodish et al. Fig. 24-17

Activation of Src
has multiple consequences

From Schwartzenberg, Oncogene 17, 1463-1468 (1998)

Recent work has provided
a more detailed model of Src activation

Closed = OFF Open = ON

Cowen-Jacob et al. Structure 13, 861-871 (2005)
Where is Src within cells?
This is a covalently attached lipid--what might that mean?
Identifying the targets of Src

Myristylation of Src is
essential for transformation
Recent work has provided
a more detailed model of Src activation

Cowen-Jacob et al. Structure 13, 861-871 (2005)

Deletion of the C-terminal region
leads to activation of v-Src

Lodish et al. Fig. 24-17

c- Src is a tyrosine kinase

What does it do in the cell?

What are its targets?

Figure 5.12b The Biology of Cancer (© Garland Science 2007)
Figure 5.12a The Biology of Cancer (© Garland Science 2007)
Figure 5.11 The Biology of Cancer (© Garland Science 2007)